A computer-implemented method, computer program product, control device, and system for controlling a sample preparation system

Abstract

A computer-implemented method for controlling a sample preparation system (1) for preparing chemical, pharmaceutical, and / or biotechnological samples is provided. The sample preparation system (1) comprises a fluid path and at least one consumable (2a), such as a filter, provided on the fluid path. The method includes controlling the sample preparation system (1) such that a sample fluid containing a target substance flows through the fluid path through at least one consumable (2a), obtaining system information (step S14), and determining a regression function using the system information obtained for a specified period preceding the current time (t current ), determining whether a first specified condition is satisfied, the first specified condition being determined to be satisfied when a value indicating a predicted blockage amount (Vpb) corresponds to the amount of sample fluid (Vin_t current ) introduced into the fluid path at the current time (t current ), the predicted blockage amount (Vpb) being the value of the independent variable (x) of the regression function when the value of the dependent variable (y) is a specified maximum differential pressure, (step S18), if the first specified condition is satisfied, controlling the sample preparation system (1) to stop introducing the sample fluid into the fluid path (step S20), if the first specified condition is not satisfied, controlling the sample preparation system (1) to continue introducing the sample fluid into the fluid path, obtaining system information (step S14), determining a regression function (step S16), and determining whether the first specified condition is satisfied (step S18), and repeating the steps.

Description

Technical Field

[0001] The present application relates to a computer-implemented method, a computer program product, a control device, and a system for controlling a sample preparation system.

Background Art

[0002] In order to generate a target substance such as a protein by purification, a sample containing the target substance is usually subjected to a clarification step before purification to remove contaminants such as whole cells, cell fragments, host cell nucleic acids, and host cell proteins derived from host cells, contaminants derived from the medium such as proteinaceous and non-proteinaceous medium components, and contaminants derived from vectors including vector nucleic acids and viral vectors. In the clarification step, for example, the sample may be filtered through a filter.

[0003] In order to obtain a high yield of the target substance, it would be desirable to leave as little of the target substance as possible within a sample preparation system configured to process the sample (e.g., within a filter used as the sample preparation system, within a fluid path included in the sample preparation system, etc.).

Summary of the Invention

[0004] According to one aspect, the problem relates to improving the efficiency of generating a target substance contained in a sample. This problem is solved by the features disclosed in the independent claims. Further exemplary embodiments are defined by the dependent claims.

[0005] According to one aspect, a computer-implemented method for controlling a sample preparation system for preparing chemical, pharmaceutical, and / or biotechnological samples is provided, the sample preparation system comprising a fluid path and at least one consumable such as a filter provided on the fluid path. The method comprises Controlling a sample preparation system such that a sample fluid containing a target substance flows through at least one consumable within a fluid path, wherein the sample fluid can pass through an upstream point within the fluid path, the at least one consumable, and a downstream point within the fluid path in this order, the step; at least the amount of sample fluid introduced into the fluid path, the upstream pressure at the upstream point, the downstream pressure at the downstream point, and obtaining system information indicating the above; Determining a regression function using the system information obtained for a specified period (Δt) preceding the current time (t current ), wherein the regression function is such that at different times (t i -Δt,…,t i ) within the specified period (Δt), the differential pressure (dP_t i -Δt,…,dP_t i ) across at least one consumable, based on the upstream pressure and the downstream pressure, obtained for these times, is the value of the dependent variable (y) of the regression function, and at different times (t i -Δt,…,t i ) within the specified period (Δt), the amount of sample fluid (Vin_t i -Δt,…,Vin_t i ) introduced into the fluid path, obtained for these times, is the value of the independent variable (x) of the regression function, and is determined by making this assumption, the step; Determining whether a first specified condition is satisfied, wherein the first specified condition is satisfied and determined when a value indicating a predicted blockage amount (Vpb) corresponds to the amount of sample fluid (Vin_t current ) introduced into the fluid path at the current time (t current ), and the predicted blockage amount (Vpb) is the value of the independent variable (x) of the regression function when the value of the dependent variable (y) is the specified maximum differential pressure, the step; If the first specified condition is satisfied, controlling the sample preparation system to stop introducing the sample fluid into the fluid path, the step; If the first specified condition is not satisfied, controlling the sample preparation system to continue introducing the sample fluid into the fluid path; repeating the steps of obtaining system information, determining a regression function, and determining whether the first specified condition is satisfied; and including.

[0006] In various aspects and embodiments described herein, a "fluid path" can be formed, for example, by at least one or more tubes. In some exemplary embodiments, the (one or more) tubes may be selected from plastic polyvinyl chloride, polypropylene, polyethylene, polytetrafluoroethylene, and fluorinated ethylene propylene. Thereby, the (one or more) tubes can provide flexibility and chemical resistance.

[0007] In various aspects and embodiments described herein, the "at least one consumable" may be a filter. More specifically, the "at least one consumable" may be a clarification filter for clarifying a sample fluid containing a target substance. In some exemplary embodiments, the "clarification filter" may be a depth filter and / or a membrane, preferably a depth filter. Further, in some situations, the "clarification filter" may be a cell culture fluid clarification filter configured to separate cells and other contaminants from a sample.

[0008] In various aspects and embodiments described herein, the "at least one consumable" may be a disposable consumable. As used herein, the term "disposable consumable" means that the consumable cannot be cost-effectively recovered after use, for example due to blockage, and is discarded.

[0009] In the various aspects and embodiments described herein, the "target substance" is not specifically limited. For example, the target substance may be any biological molecule for any purpose, such as an antibody, a hormone, a vaccine, a nucleic acid, an exosome and a virus, and a protein such as a virus-like particle. In some exemplary embodiments, the target substance may be an antibody, more specifically a monoclonal antibody (mAb), or a fragment or derivative thereof, or a nanobody. Examples of monoclonal antibodies may include, but are not limited to, adalimumab, cetuximab, rituximab, infliximab, omalizumab, and denosumab. The target substance can be obtained from mammalian cells such as, for example, "Chinese hamster ovary cells" (CHO cells), HeLa, or human umbilical vein endothelial cells (HUVEC), bacterial cells, or insect cells, media, and cell lines.

[0010] In the various aspects and embodiments described herein, the "sample fluid" contains a plurality of components and is not particularly limited as long as at least one of the plurality of components of the sample fluid is one of the above-described target substances. Further (impurity) components (contaminants) are not particularly limited and may depend on the preparation conditions of the target substance. Examples of further components may include, but are not limited to, aggregates, host cell proteins, deoxyribonucleic acids, and fragments and charge variants thereof. The sample fluid can be obtained by applying any biological, biochemical, chemical, or pharmaceutical method. Thereby, the sample fluid can be obtained by performing in advance a purification method that applies different purification units. For example, the target substance can be produced by an appropriate cell line such as a CHO cell line, for example, by perfusion culture.

[0011] In various aspects and embodiments described herein, at least a portion of the "system information" can be obtained from sensors disposed at positions suitable for collecting respective system information for the sample preparation system. Further, in various aspects and embodiments described herein, at least a portion of the "system information" may be obtained by accessing a storage medium that stores the relevant (one or more) system information. The storage medium may be either internal or external to a control device configured to execute a method according to any one of the various aspects and embodiments described herein.

[0012] For example, system information indicating the amount of sample fluid introduced into a fluid path can be obtained from a rotational speed sensor provided on a pump configured to move the sample fluid within the fluid path, and the rotational speed sensor can measure the rotational speed of the pump. Since the amount of sample fluid introduced into the fluid path can be proportional to the amount of rotation of the pump, the amount of sample fluid can be calculated from the measured rotational speed of the pump (e.g., by multiplying the rotational speed by the time elapsed since the start of the sample preparation process executed by the sample preparation system). In various aspects and embodiments described herein, the "amount" of a fluid (e.g., sample fluid) can be understood as the volume of the fluid. In some exemplary embodiments, the amount of rotation of the pump for calculating the amount of sample fluid introduced into the fluid path can be obtained without a rotational speed sensor. For example, a calibrated value of the rotation of the pump (driven, e.g., by a DC motor or a stepper motor) may be obtained from the storage medium as described above, and the amount of sample fluid can be calculated using the obtained calibrated value of the rotation of the pump.

[0013] Furthermore, for example, system information indicating upstream pressure and downstream pressure can be obtained from a pressure sensor provided at the upstream point and a pressure sensor provided at the downstream point, respectively. However, regarding the downstream pressure, in some exemplary embodiments, it may be atmospheric pressure instead of the pressure measured by the pressure sensor provided at the downstream point. In such exemplary embodiments, the storage medium as described above may store the value of atmospheric pressure, and the value of atmospheric pressure as the downstream pressure may be obtained from the storage medium, for example.

[0014] In various aspects and embodiments described herein, the "designated period" Δt can be specified by a unit of time (e.g., seconds, minutes, etc.) and / or by a change in the amount (e.g., volume) of fluid introduced into the fluid path. When specifying the "designated period" by a change in the amount of fluid introduced into the fluid path, the "designated period" of x ml may be expressed as Δt = x ml, and may indicate the period during which x ml of fluid is introduced into the fluid path. The amount of fluid introduced into the fluid path can be calculated, for example, from the rotational speed of the pump as described above. It should be noted that when the "designated period" is specified by a change in the amount of fluid introduced into the fluid path, the actual time elapsed within the amount of fluid introduced into the fluid path for the same amount of fluid can be different in different iterations of the steps of acquiring system information, determining a regression function, and determining whether a first specified condition is satisfied.

[0015] Furthermore, in various aspects and embodiments described herein, the "current time" t current and "different times", for example t i -Δt,…,t i etc. (i = 0, 1, 2, …) can each be represented by a unit of time (e.g., x seconds, minutes, etc. from the start of a sample preparation process performed by a sample preparation system) and / or by the amount (e.g., volume) of fluid introduced into the fluid path from the start of the sample preparation process to the relevant time point. Thus, in the present disclosure, the "current time" t current can also be understood as the current amount of fluid introduced into the fluid path. More generally, the time point t iIt may also be understood as the amount of fluid introduced into the fluid path up to that point in time.

[0016] In some exemplary embodiments, the system information "acquired with respect to" a specified period Δt preceding the current time t may include one or more system information acquired during the specified period Δt. Additionally, in some exemplary embodiments, the system information "acquired with respect to" the specified period Δt may further include one or more system information acquired outside the specified period Δt but considered applicable to the specified period Δt. For example, if the atmospheric pressure is acquired as the downstream pressure without a pressure sensor at the downstream point, the atmospheric pressure does not necessarily have to be "acquired during" the specified period Δt, but the acquired atmospheric pressure may be considered as the downstream pressure during the specified period Δt. current

[0017] In various aspects and embodiments described herein, the "differential pressure across at least one consumable" at a particular time t i can be obtained by calculating the difference between the downstream pressure at a particular time t i and the upstream pressure at a particular time t. i

[0018] In various aspects and embodiments described herein, the specified period Δt preceding the current time t may include a discrete number of time points. The discrete number of time points may be correlated or dependent on the sampling rate of the participating sensors such as pressure sensors. Further, the specified period Δt preceding the current time t can indicate the period from time t current -Δt to time t current and t i may be a time point before the current time t i or before the current time t i current current current In some exemplary embodiments, t i i is before the current time t, in other words, it may be a single point immediately before the current time t current current current i i is more than a single point before the current time tcurrent It may be further away, preferably within a specific ratio (e.g., 1 / 10) of the total expected time of the sample preparation process (or the total amount of the sample fluid).

[0019] In some exemplary embodiments, the "specified period", "specified maximum differential pressure", and "first specified condition" may be specified by the user using an input device.

[0020] In some exemplary embodiments, the "first specified condition" may be determined to be satisfied when the difference between the "value indicating the predicted occlusion amount (Vpb)" and the "amount of the sample fluid introduced into the fluid path at the current time (Vin_t current )" is equal to or less than a specified threshold value (in other words, the value indicating the predicted occlusion amount (Vpb) may be regarded as corresponding to the amount of the sample fluid introduced into the fluid path at the current time (Vin_t current )). On the other hand, the "first specified condition" may be determined not to be satisfied when the difference is greater than the "specified threshold value" (in other words, the value indicating the predicted occlusion amount (Vpb) may not be regarded as corresponding to the amount of the sample fluid (Vin_t current )).

[0021] In various aspects and embodiments described herein, the "amount of the sample fluid introduced into the fluid path at the current time (t current ) (Vin_t current )" may be understood as the amount of the sample fluid currently introduced into the fluid path.

[0022] In various aspects and embodiments described herein, the "predicted occlusion amount" can be regarded as the amount of sample fluid that can block the flow through at least one consumable (e.g., a filter) when introduced into at least one consumable. Thus, by controlling the sample preparation system to stop introducing the sample fluid into the fluid path when a specified threshold is met as in the method according to the above aspect, the amount of sample fluid introduced into the fluid path but unable to pass through at least one consumable can be reduced, in other words, the amount of sample fluid that can be wasted can be reduced. This can contribute to improving the efficiency of generating the target substance contained in the sample fluid.

[0023] In some exemplary embodiments, the method according to the above aspect further comprises the step of further controlling the sample preparation system such that when a first specified condition is met, the rinse fluid flows through at least one consumable within the fluid path and may further be included.

[0024] Due to the flow of the rinse fluid, the sample fluid still present within the fluid path upstream of at least one consumable and / or the sample fluid still contained within at least one consumable can be pushed to pass through at least one consumable. This can contribute to increasing the yield of the target substance in the sample fluid.

[0025] In some exemplary embodiments, the method according to the above aspect further comprises the step of further controlling the sample preparation system such that when a first specified condition is met, air flows through at least one consumable within the fluid path and may further be included.

[0026] The sample fluid that still exists within the fluid path before reaching at least one consumable due to the air flow (in other words, the sample fluid that still exists within the fluid path upstream of at least one consumable), and / or the sample fluid that is still contained within at least one consumable, can be pushed to pass through at least one consumable. This can contribute to increasing the yield of the target substance in the sample fluid.

[0027] In some exemplary embodiments, the method according to the above-described aspect further comprises controlling the sample preparation system to remove at least one consumable from the fluid path when a first specified condition is satisfied. may further include.

[0028] Also, the method according to the above-described aspect further comprises controlling the sample preparation system to provide a new consumable on the fluid path after further controlling the sample preparation system to remove at least one consumable from the fluid path. may further include.

[0029] The control of the sample preparation system to provide a new consumable on the fluid path can be executed when a second specified condition is satisfied. The second specified condition may include a condition regarding the amount of the remaining sample fluid to be processed by the sample preparation system. For example, a threshold value of the amount of the remaining sample fluid may be specified (e.g., by the user), and the second specified condition is determined to be satisfied when the amount of the remaining sample fluid exceeds the threshold value. Further, in some exemplary embodiments, the threshold value of the amount of the remaining sample may be determined by a control device configured to control the sample preparation system.

[0030] In some exemplary embodiments, at least one consumable may be a filter. In such an exemplary embodiment, the sample preparation system receives at least one consumable provided on the fluid path, provides at least one consumable on the fluid path, It may further include a filter handling device configured as described above.

[0031] Furthermore, the step of controlling the sample preparation system to remove at least one consumable from the fluid path may include the step of outputting a control signal for removing at least one consumable from the fluid path to the filter handling device by receiving the at least one consumable. Further, the step of controlling the sample preparation system to provide a new consumable on the fluid path may include the step of outputting a control signal for providing a new consumable on the fluid path to the filter handling device.

[0032] In some exemplary embodiments, the sample preparation system may further include a pump configured to move fluid within the fluid path. The pump may be, for example, a peristaltic pump, a diaphragm pump, or a piston pump. In an exemplary embodiment where the sample preparation system further includes a pump, the step of controlling the sample preparation system such that the sample fluid flows through at least one consumable within the fluid path may include the step of outputting a control signal for moving the sample fluid within the fluid path towards the at least one consumable to the pump. Further, the step of controlling the sample preparation system to stop introducing the sample fluid into the fluid path may include the step of outputting a control signal for stopping the movement of the sample fluid within the fluid path to the pump.

[0033] In some exemplary embodiments, the sample preparation system may include at least an open state in which the sample fluid can flow towards at least one consumable within the fluid path, or a closed state in which the flow of the sample fluid towards at least one consumable within the fluid path is prevented, and may further include a valve configured to switch between them.

[0034] In an exemplary embodiment where the sample preparation system further comprises a valve as described above, the step of controlling the sample preparation system such that the sample fluid flows through at least one consumable within the fluid path may include outputting a control signal to the valve to set the valve to an open state. Further, the step of controlling the sample preparation system to stop introducing the sample fluid into the fluid path may include outputting a control signal to the valve to set the valve to a closed state.

[0035] In some exemplary embodiments, the valve may be a three-way valve (e.g., a three-way diaphragm valve or a three-way ball valve) having three ports (e.g., a first, a second, and a third port), and capable of switching between three states: a closed state in which fluid flow through the valve is prevented, a first open state in which fluid can flow from the first port to the second port (or vice versa), and a second open state in which fluid can flow from the first port to the third port (or vice versa). In such an exemplary embodiment, the first port of the three-way valve may be connected to a source of the sample fluid, and either the second port or the third port of the three-way valve may be connected to, for example, a part of the fluid path leading to at least one consumable. The remaining port of the three-way valve may be connected to, for example, another part of the fluid path leading to a waste container of the sample preparation system.

[0036] In some exemplary embodiments, the step of determining the regression function is obtaining one or more values of one or more parameters related to the sample fluid and / or the sample preparation system, and for each of the one or more parameters, generating an equation for determining the occlusion amount as a function of one of the one or more parameters, wherein the occlusion amount indicates the amount of the sample fluid that can block the flow of the sample fluid through at least one consumable when introduced into the at least one consumable, for each of the one or more parameters, the amount of the sample fluid introduced into the fluid path at the current time (t current ) (Vin_t currentor the total amount of sample fluid introduced into the fluid path, the amount of occlusion obtained using the generated formula, determining a coefficient indicating the ratio between; determining a regression function using the coefficient determined for each of the one or more parameters; may include. In such an exemplary embodiment, the one or more parameters are the cell density of the sample fluid, the turbidity of the sample fluid, the particle size distribution of the sample fluid, the optical density of the sample fluid, the filter resistance index when at least one consumable is a filter, the filter surface area when at least one consumable is a filter, may include one or more of.

[0037] In some exemplary embodiments, one or more values of the one or more parameters may be stored in a storage medium (e.g., internal or external to a control device configured to control a sample preparation system) as described above and retrieved therefrom.

[0038] Furthermore, the step of determining a regression function using the coefficient determined for each of the one or more parameters includes determining a sensitivity coefficient using the coefficient determined for each of the one or more parameters; selecting, based on the sensitivity coefficient, one type of regression function to be used as the regression function from among a plurality of types of regression functions; may include.

[0039] In various aspects and embodiments described herein, the sample preparation system may be for preparing a sample that is a cell culture fluid containing protein. In such a case, the sample fluid may be a cell culture fluid, and the protein contained in the cell culture fluid may be the target substance.

[0040] According to another aspect, a computer program product is provided. According to another aspect, a computer program product is provided. The computer program product includes computer-readable instructions that, when loaded and executed on a suitable system, cause the system to execute a method according to any one of the above aspects and its exemplary embodiments.

[0041] According to yet another aspect, a control device is provided. The control device includes a processor configured to execute a method according to any one of the above aspects and its exemplary embodiments, and a storage medium communicating with the processor. It is provided with.

[0042] According to yet another aspect, a system is provided. The system is a sample preparation system, including a fluid path, and at least one consumable such as a filter provided on the fluid path, and a sample preparation system provided with. and a control device according to the above aspect. It is provided with.

[0043] The various aspects and embodiments described herein can be applied to single-pass filtration of biopharmaceutical cell suspensions, for example, to the small-scale clarification of mAb-expressing mammalian suspension cell cultures. Since the cell suspension has a high particulate load and a clarification process over seconds or minutes can already cause filter blockage, pressure changes in the last seconds or minutes (or the last few ml depending on the scale) can be important in predicting filter blockage. In the various aspects and embodiments described herein, by setting the "designated period" to seconds or minutes, for example, it is possible to enable prediction of filter blockage in such a clarification process. Further, in such a clarification process, the volume of the cell suspension in the (depth) filter and the supply line can be a significant proportion of the volume being filtered. Further, the volume of the cell suspension in the (depth) filter and the supply line can be important in causing filter blockage. The various aspects and embodiments described herein can take into account such volumes (as the amount of sample fluid introduced into the fluid path) to predict filter blockage.

[0044] Also, in single-pass filtration of biopharmaceutical cell suspensions, the value of the product (e.g., the target substance) in the filter (e.g., depth filter) can be important. Even without blockage occurring, a small amount (e.g., 5 to 20%) of the product can always be trapped in each filter at the end of filtration. Thus, accurate prediction of blockage can be important. If the prediction is too conservative, more filters are used and as a result, more product loss occurs. If the prediction is not conservative enough, blockage is more likely to result in more product loss (e.g., perhaps 50% product loss).

[0045] Further, the various aspects and embodiments described herein can follow the input fluid with an (user-definable) amount of uncontaminated rinse fluid to achieve maximum product recovery.

[0046] The various aspects and embodiments described herein may be compatible with simultaneous clarification and purification processes, such as those used on sample preparation systems.

[0047] The various aspects and embodiments described herein may be used for any clarification application where an appropriate control system is available.

[0048] Furthermore, the various aspects and embodiments described herein may be used for any application where a dead-end filter that can be blocked is used and an optimal recovery rate is desired.

[0049] Furthermore, the various aspects and embodiments described herein may be applicable to microbial cultures, cultures of different scales, and as a method or system for fractionating an output.

[0050] Since the various aspects and embodiments described herein do not depend on historical data for determining a regression function, any one of the various aspects and embodiments described herein can provide flexible and robust real-time control of a sample preparation system, which can be ideal for research and development and early process development. The variability between (especially small-scale) filters and between actual culture lots themselves can reduce the influence of a given model. The adaptability of the various aspects and embodiments described herein can accommodate such variability.

[0051] Any one of the various aspects and embodiments described herein can provide one or more of the following technical effects. - Achieve an optimal ratio of output volume to contaminant concentration, - Obtain maximum product recovery from consumables (e.g., filters), which may be completely blocked before the end of rinsing, - Optimize the efficiency of the clarification process to supply the maximum volume and quality of output samples with the minimum possible time requirements and maximum sample / time / cost, -Fully automate the sample preparation process and reduce the time requirements for operators.

[0052] The subject matter described in this application can be implemented, in some cases, in the form of one or more computer program products, as a method or a system. The subject matter described in this application can be implemented in a data signal or a machine-readable medium, and the medium is embodied in one or more information carriers such as a CD-ROM, a DVD-ROM, a semiconductor memory, or a hard disk. Such a computer program product can cause a data processing device to execute one or more operations described in this application.

[0053] Furthermore, the subject matter described in this application can also be implemented as a system including a processor and a memory coupled to the processor. The memory can encode one or more programs to cause the processor to execute one or more of the methods described in this application. In some examples, the system may be a general-purpose computer system. In other examples, the system may be a dedicated computer system including an embedded system.

Brief Description of the Drawings

[0054] Details of one or more implementations are described in the following exemplary drawings and explanations. Other features will become apparent from the description, the drawings, and the claims. However, it should be understood that even if the embodiments are described separately, a single feature of different embodiments can be combined in a further embodiment.

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DETAILED DESCRIPTION OF THE INVENTION

[0055] Detailed Description of Embodiments Hereinafter, examples will be described in detail with reference to the drawings. It should be understood that various changes can be made to the examples. In particular, one or more elements of one example can be combined and used in other examples to form new examples.

[0056] Sample Preparation System In the present disclosure, the sample preparation system can be understood, for example, as a system for preparing chemical, pharmaceutical, and / or biotechnological samples to generate target substances contained in the samples. For example, the sample may be a cell culture solution containing a protein as a target substance.

[0057] In some exemplary embodiments, the sample preparation system can be configured to perform one or more unit operations such as clarification and purification on the sample. For each of the one or more unit operations, a consumable such as a filter may be provided on the sample preparation system. For example, a clarification filter may be provided for clarification. Further, for example, a purification filter or a purification chromatography column may be provided for purification. More specifically, the sample preparation system may include a fluid path, and a sample fluid containing a target substance can flow through one or more consumables within the fluid path and be subjected to one or more unit operations, and one or more consumables for the one or more unit operations may be provided on the fluid path.

[0058] Hereinafter, as shown in FIG. 1, exemplary embodiments will be described with respect to an exemplary sample preparation system configured to perform clarification and / or purification on a sample. However, it should be noted that the various aspects and embodiments described herein can also be used to control a sample preparation system having a configuration different from the exemplary sample preparation system shown in FIG. 1.

[0059] FIG. 1 shows an exemplary sample preparation system configured to perform clarification and / or purification on a sample. As shown in FIG. 1, the sample preparation system 1 may comprise a fluid path including an input line 12, a first line 6, a second line 7, a third line 8, a fourth line 19, an output line 22, and / or a waste container line 44. Some or all of the lines 12, 6, 7, 8, 19, 22, and 44 may be made of a chemically resistant flexible tube. In the sample preparation system 1, a fluid (e.g., a sample fluid containing a target substance) may flow in the direction indicated by the arrow shown in FIG. 1 within the fluid path. Hereinafter, the term "upstream" with respect to a reference point within the fluid path may be understood as any point in a part of the fluid path through which the fluid can pass from the fluid source to the reference point when the fluid flows. Further, the term "downstream" with respect to the reference point may be understood as any point in a part of the fluid path through which the fluid can pass after the fluid has passed through the reference point when the fluid flows.

[0060] Furthermore, the sample preparation system 1 may comprise a series arrangement of a first means 2 configured to be connected to a first consumable 2a and a second means 3 configured to be connected to a second consumable 3a. In the specific example of FIG. 1, the first consumable 2a is a clarification filter, and the second consumable 3a is a purification filter or a purification chromatography column. The clarification filter may be a depth filter and / or a membrane.

[0061] For example, the purification filter may be a membrane, and the purification chromatography column may contain functionalized beads. The second consumable 3a may be configured to bind and elute the target substance. The second means 3 may be disposed downstream of the first means 2 within the sample preparation system 1, and the first means 2 and the second means 3 may be integrally connected to each other within the sample preparation system 1. As used herein, the term "integrally connected" means being disposed within one device (e.g., within the sample preparation system 1) and not being disposed within separate devices.

[0062] The sample preparation system 1 can be configured to perform, in an automated manner within a single device, a combination of clarification as a first unit operation and purification as a second unit operation. Thus, each method can be faster and the process efficiency can be significantly improved compared to methods performed within two separate devices.

[0063] In the sample preparation system 1, the sample fluid can be introduced from an input cup 13 configured to hold the fluid to be prepared by the sample preparation system 1, e.g., the sample fluid, into an input line 12 disposed upstream of the first means 2. In some exemplary embodiments, the input cup can have a volume of from 10 ml to 150 ml, preferably from 20 ml to 80 ml. The input line 12 can provide fluid communication between the first means 2 and the input cup 13.

[0064] Furthermore, in the sample preparation system 1, an input sample valve 14 may be provided downstream of the input cup 13 on the input line 12 and upstream of the first means 2. Thus, the input sample valve 14 can divide the input line 12 into a portion upstream of the input sample valve 14 and a portion downstream of the input sample valve 14. The input sample valve 14 can be configured to direct fluid to the input line 12 downstream of the input sample valve 14 or to a waste container (not shown). For example, when the fluid is directed to the input line 12 downstream of the input sample valve 14, the flow of fluid to the waste container can be blocked. Further, for example, when the fluid is directed to the waste container, the flow of fluid to the input line 12 downstream of the input sample valve can be blocked. The input sample valve 14 can be a three-way valve (e.g., a three-way diaphragm valve or a three-way ball valve).

[0065] Furthermore, the sample preparation system 1 may include a process fluid input valve 24 disposed upstream of the first means 2. Each of the process fluid input valves 24 may be a two-way valve that can be switched between an open state that allows fluid to flow through the valve and a closed state that prevents fluid from flowing through the valve. Various types of process fluids may be stored, for example, in bottles and connected by lines to one of the process fluid input valves 24. Each of the process fluid input valves 24 may permit or stop the flow of the corresponding type of process fluid into the input line 12.

[0066] The different types of process fluids that may flow into the input line 12 via the process fluid input valve 24 may include, for example, fluids used to wash the sample preparation system 1 and / or fluids used in clarification and / or purification. Examples of process fluids used to wash the sample preparation system 1 may include, but are not limited to, buffer solutions, sodium hydroxide, and / or water. In the specific example shown in FIG. 1, the process fluid input valve 24 includes at least valves for permitting or stopping the flow of rinse fluid and air.

[0067] Further examples of process fluids may include, but are not limited to, phosphate buffered saline (PBS), elution buffer, low pH strip buffer, etc. The elution buffer is not particularly limited, and its selection may depend on the substance to be eluted. The elution buffer may be selected, for example, from citrate buffer, glycine / HCl buffer, and acetate buffer.

[0068] Furthermore, the sample preparation system 1 may include a pump 9 provided on the input line 12 downstream of the input sample valve 14 and the process fluid input valve 24. Further, the pump 9 may be provided upstream of the first means 2. The pump 9 may be configured to move fluid within the fluid path of the sample preparation system 1. In the specific example of FIG. 1, the pump 9 is a peristaltic pump. In other exemplary embodiments, the pump 9 may be a diaphragm pump or a piston pump.

[0069] The input line 12 can provide fluid communication between the first means 2 and one or more of the input sample valve 14 and the process fluid input valve 24.

[0070] In the specific example of FIG. 1, the first means 2 is a first filter holder including a first clamp 10a configured to clamp a first consumable 2a (e.g., a clarification filter) on an upper surface (in other words, an upstream surface), and a second clamp 10b configured to clamp the first consumable 2a on a lower surface (in other words, a downstream surface). Thereby, the first consumable 2a can be firmly supported against the internal pressure during operation. Each of the first clamp 10a and the second clamp 10b may include an O-ring configured to connect the upstream and downstream fluid paths of the first means 2 when the O-ring is in a compressed state (in other words, when the clamp is closed). In some exemplary embodiments, one of the first clamp 10a and the second clamp 10b may be a fixed clamp (in other words, this does not move), and the other may be a movable clamp. Thereby, the first consumable 2a can be clamped by pressing the movable clamp against the fixed clamp. Preferably, the first clamp 10a may be a fixed clamp, and the second clamp 10b may be a movable clamp.

[0071] In some exemplary embodiments, the first means 2 does not necessarily have to be a first filter holder having the first and second clamps 10a, 10b as described above. For example, the first means 2 may be an upstream end of the first line 6 that supports the first consumable 2a from the downstream side. When the first consumable 2a is supported from its downstream side by the upstream end of the first line 6, the first consumable 2a may be supported from its upstream side by the downstream end of the input line 12 connected to the first means 2.

[0072] Furthermore, the sample preparation system 1 may include an input stack 11 configured to hold at least one first consumable 2a. In some exemplary embodiments, the input stack 11 may be configured to hold from 1 to 150 first consumables 2a, preferably from 1 to 50 first consumables 2a. The sample preparation system 1 may further include a filter handling device (not shown in FIG. 1) connectable to the first means 2. The filter handling device may be configured to provide the first consumable 2a to the first means 2 and receive the first consumable 2a from the first means 2. Details of the filter handling device will be described later with reference to FIGS. 2 and 3.

[0073] As shown in FIG. 1, the first line 6 may provide fluid communication between the first means 2 and a first valve 4 disposed between the first means 2 and the second means 3. The first valve 4 may be provided on the fluid path between the first means 2 and the second means 3. The first valve 4 may be configured to direct fluid from the first line 6 of the sample preparation system 1 to the second line 7 or the third line 8. For example, when the fluid is directed to the second line 7, the flow of fluid to the third line 8 may be blocked. Further, for example, when the fluid is directed to the third line 8, the flow of fluid to the second line 7 may be blocked. The first valve 4 may be, for example, a three-way diaphragm valve or a three-way ball valve.

[0074] As can be seen from FIG. 1, the second line 7 may provide fluid communication between the first valve 4 and a second valve 5 downstream of the second means 3 via the second means 3. Further, the third line 8 may provide direct fluid communication between the first valve and the second valve 5. Thus, the third line 8 may be configured to bypass the second means 3.

[0075] The second means 3 may be connected to a second consumable 3a (e.g., a purification filter or a purification chromatography column) and configured to support the second consumable 3a. For example, the second means 3 may be a second filter holder or may correspond to a portion of the second line 7 that provides fluid communication between the first valve 4 and the second valve 5.

[0076] When the second means 3 is the second filter holder, the second line 7 may be interrupted between the first valve 4 and the second valve 5. The second filter holder as the second means 3 is disposed at the interruption location and may be connected to the first end of the second line 7 interrupted on the upstream side and the second end of the second line 7 interrupted on the downstream side. In some exemplary embodiments, the second filter holder as the second means 3 may include a first clamp and a second clamp configured to clamp the second consumable 3a in a manner similar to the first clamp 10a and the second clamp 10b of the first means 2 as described above.

[0077] When the second means 3 corresponds to a part of the second line 7, the second line 7 may be interrupted between the first valve 4 and the second valve 5. The second consumable 3a may be disposed at the interruption location and supported by the first end of the second line 7 interrupted on the upstream side and the second end of the second line 7 interrupted on the downstream side.

[0078] The second valve 5 disposed downstream of the second means 3 may be configured to direct fluid to a fourth line 19 downstream of the second valve 5. For example, the second valve 5 may have three ports with two inlets connected to the second line 7 and the third line 8, and may be a three-way valve that can be in one of two states that direct flow from one of the two inlets, e.g., the second line 7 and the third line 8, to the fourth line 19 downstream of the second valve 5. In some exemplary embodiments, the second valve may be a three-way diaphragm valve or a three-way ball valve.

[0079] The fourth line 19 may provide fluid communication between the second valve 5 and an output valve 20 disposed downstream of the second valve 5. The output valve 20 may be configured to direct fluid to an output line 22 that provides fluid communication between the output valve 20 and an output cup 23, or to a waste container line 44 that provides fluid communication between the output valve 20 and a waste container (not shown). For example, when fluid is directed to the output line 22, the flow of fluid to the waste container line 44 may be blocked. Further, for example, when fluid is directed to the waste container line 44, the flow of fluid to the output line 22 may be blocked. The output valve 20 may be a three-way valve having three ports and may be in one of two states that direct flow to one of two outlets, such as the output line 22 and the waste container line 44. In some exemplary embodiments, the output valve 20 may be a three-way diaphragm valve or a three-way ball valve.

[0080] The output cup 23 may be configured to hold fluid output by the sample preparation system 1. In some exemplary embodiments, the output cup may have a volume from 10 ml to 150 ml, preferably from 20 ml to 80 ml.

[0081] Furthermore, the sample preparation system 1 shown in FIG. 1 may include a waste pump 25 configured to pump fluid in the fluid path to direct the fluid to a waste container (not shown). For example, fluid directed from the output valve 20 to the waste container line 44 and / or fluid flowing from the input sample valve 14 (when the input sample valve 14 is set to prevent the sample fluid from flowing toward the first means) may be directed to the waste container by operating the waste pump 25.

[0082] Several sensors may be provided on the sample preparation system 1 shown in FIG. 1.

[0083] For example, a first liquid sensor 15 may be provided on the input line 12 at a position downstream of the input sample valve 14 and upstream of the first means 2. The first liquid sensor 15 may be configured to detect whether or not there is liquid at the position where the first liquid sensor 15 is disposed. According to this configuration, the first liquid sensor 15 can detect, for example, whether or not the input cup 13 is empty.

[0084] Furthermore, for example, a second liquid sensor 16 may be provided on the fluid path at a position downstream of the first means 2 and upstream of the second means 3. The second liquid sensor 16 may be configured to detect whether or not there is liquid at the position where the second liquid sensor is disposed. Preferably, the second liquid sensor 16 may be disposed on the first line 6 upstream of the first valve 4. According to this configuration, based on the detection result of the second liquid sensor 16, the first valve 4 and the second valve 5 can be switched, for example, to direct the fluid through the second line 7 via the second means 3 or through the third line 8 to bypass the second means 3.

[0085] Furthermore, for example, a first pressure sensor 17 may be provided on the input line 12 at a position downstream of the first liquid sensor 15 and upstream of the first means 2, in other words, at an upstream point with respect to the first means 2. The first pressure sensor 17 may be configured to detect the pressure of the sample preparation system 1 at the position where the first pressure sensor 17 is disposed. Preferably, the first pressure sensor 17 may be disposed downstream of the pump 9 and upstream of the first means 3. In some exemplary embodiments, the first pressure sensor 17 may be disposed downstream of the pump 9 and the first liquid sensor 15 and upstream of the first means 9. Thereby, the flow of fluid to the first means can be easily controlled.

[0086] Furthermore, for example, a second pressure sensor 18 may be provided on the fluid path downstream of the first means 2 and upstream of the second means 3, in other words, at a downstream point with respect to the first means 2. The second pressure sensor 18 may be configured to detect the pressure of the sample preparation system 1 at the position where the second pressure sensor 18 is disposed. Preferably, the second pressure sensor may be disposed on the first line 6 upstream of the first valve 4. According to this configuration, based on the measurement result of the second pressure sensor 18, the first valve 4 and the second valve 5 can be switched to direct the fluid through the second line 7 via the second means 3 or through the third line 8 to bypass the second means 8.

[0087] Furthermore, for example, a target substance detection sensor 21 may be provided on the fourth line 19 at a position downstream of the second valve 5 and upstream of the output valve 20, i.e., upstream of the fourth line 19. The target substance detection sensor 21 may be configured to detect the target substance in the fluid path at the position where the target substance detection sensor 21 is disposed. According to this configuration, the target substance coming out of the first means 2 and / or the second means 3 can be easily detected. Depending on the detection result of the target substance detection sensor 21, the fluid can be directed to the output cup 23 or the waste container. Specifically, when the target substance is detected by the target substance detection sensor 21, the output valve 20 can be switched to direct the target substance or the fluid containing the target substance to the output cup 23. When the target substance is not detected by the target substance detection sensor 21, the output valve 20 can be switched to direct the fluid to the waste container. In some exemplary embodiments, the target substance detection sensor 21 may be a UV sensor.

[0088] FIG. 2 shows a side view of a part of an exemplary sample preparation system. Specifically, FIGS. 2(a) and 2(b) show an input stack 11 that holds a plurality of filters as a first consumable 2a (e.g., a clarification filter), a filter handling device 27, and a waste bin 26. The filter handling device 27 can be connected to the first means 2 (shown in FIG. 1) and the input stack 11. The filter handling device 27 can be configured to receive a first consumable 2a provided in a fluid path (e.g., from the first means 2) and to provide the first consumable 2a in the fluid path (e.g., to be supported by the first means 2). The filter handling device 27 can operate in an automated manner, for example, without human intervention.

[0089] Specifically, the filter handling device 27 may be connected to the input stack 11 and may be configured to receive the first consumable 2a from the first means 2 and provide the received first consumable 2a to the waste bin 26. Further, the filter handling device 27 may be configured to receive the first consumable 2a from the input stack and automatically provide the first consumable 2a to the first means 2.

[0090] In some exemplary embodiments, the filter handling device 27 may be an index wheel.

[0091] FIG. 3 shows an exemplary filter handling device in the form of an index wheel. The filter handling device 27 of FIG. 3 may have an opening 28 in its in-plane direction that conforms to the shape of a filter as the first consumable 2a at a first position of the index wheel. The filter handling device 27 shown in FIG. 3 may further have an in-plane hole 29 of the index wheel that extends from a first surface of the index wheel to a second surface of the index wheel at a second position of the index wheel. The first position of the index wheel where the opening 28 is located and the second position of the index wheel where the hole 29 is located may be diametrically opposed. Accordingly, the filter handling device 27 in the form of an index wheel may substantially have a crescent shape having a concavo-convex region delimited by two arcs.

[0092] The filter handling device 27 in the form of an index wheel can be rotated such that a first opening of a hole 29 on a first surface of the index wheel is connected to an input line 12 (see FIG. 1), and a second opening of the hole 29 on a second surface of the index wheel is connected to a first line 6 (see FIG. 1). Thus, the hole 29 on the filter handling device 27 provides fluid communication between the input line 12 and the first line 6 when the fluid needs to flow through the first means 2 without passing through the filter, for example, when only purification rather than clarification is performed by the sample preparation system 1, when the input line 12 upstream of the first means 2 is rinsed, when the sample preparation system 1 is being washed, and the like.

[0093] By rotating the filter handling device 27 in the form of an index wheel, the first consumable 2a may be received from the input stack 11 and provided to the first means 2. Also, the filter handling device 27 in the form of an index wheel may be rotated to receive the first consumable 2a from the first means and provide it to the waste bin 26.

[0094] Using the filter handling device 27 and the input stack 11 as described above with reference to FIGS. 2 and 3, the filter as the first consumable 2a can be automatically loaded into the first means 2 (e.g., the first filter holder) of the sample preparation system 1 and discharged therefrom. In other words, by using the filter handling device 27, a new (e.g., unused) first consumable 2a can be easily provided to the first means 2, while at the same time the used first consumable 2a can be easily and automatically removed.

[0095] FIG. 4 shows an exemplary pipetting robot (upper figure) and an exemplary pipette washing station (lower figure) that can be used in the sample preparation system 1. For example, the sample preparation system 1 may include a pipetting robot 30 configured to transfer a fluid such as a sample fluid from at least one input container (not shown) to the input cup 13 (see FIG. 1). The pipetting robot 30 may include a pipette tip 31 connected to a flow-through pH electrode 32 and a fluid input valve 33. The fluid input valve 33 may be configured to provide, for example, water, sodium hydroxide, and / or a pH neutralizing buffer to the pipette tip.

[0096] The pipetting robot 30 may further include a pipette liquid sensor 34 on a pipetting robot line 35 that provides fluid communication between the pipette tip 31, the flow-through pH electrode 32, and the fluid input valve 33. The pipette liquid sensor 34 may be configured to detect whether a liquid is present at its position. Also, the pipetting robot 30 may include a syringe 36 connected to the flow-through pH electrode 32. The pipette washing station 37 may be configured to receive the pipette tip 31 of the pipetting robot 30. The pipette washing station 37 may have a wall 38 that surrounds the pipette tip 31 of the pipetting robot 30 while the pipetting robot 30 is disposed therein.

[0097] Furthermore, the pipette washing station 37 may have an outlet 39 at its bottom through which fluid can exit. Specifically, the liquid can be supplied by at least one of the fluid input valves 33 of the pipetting robot 30 and can be used to wash the pipette tip 31 while the pipetting robot 30 is disposed within the pipette washing station 37. The outlet 39 may be connected to a waste container (not shown) into which the liquid can be pumped by the use of a chip washing waste pump 40. FIG. 4 further shows a storage container 41, an input container 42, and an output container 43 that may be disposed adjacent to the pipette washing station 37, for example, for storing process liquids.

[0098] In some exemplary embodiments, one input container may have a volume from 2 ml to 20 ml. The input container is not particularly limited and may be selected, for example, from culture vessels, spin tubes, and multiwell plates.

[0099] When using the pipetting robot 30, the type of input container is not particularly limited. Thus, the pipetting robot 30 can provide flexibility in the selection of the input container such that its selection can depend only on the fluid being handled, such as the sample fluid being prepared.

[0100] In addition to, or instead of, the pipetting robot 30 configured to transfer fluid from at least one input container (not shown) to the input cup 13 as described above, the sample preparation system 1 may include a pipetting robot 30 configured to transfer fluid from the output cup 23 (see FIG. 1) to at least one output container (not shown). The pipetting robot 30 configured to transfer fluid from the output cup 23 to at least one output container may be the same as the pipetting robot 30 configured to transfer fluid from at least one input container (not shown) to the input cup 13 as described above.

[0101] In some exemplary embodiments, one output container may have a volume from 2 ml to 80 ml. The output container is not particularly limited and may be selected, for example, from 15 ml centrifuge tubes, 50 ml centrifuge tubes, and multiwell plates. When using the pipetting robot 30, the type of output container is not particularly limited. Thus, the pipetting robot 30 can provide flexibility in the selection of the output container.

[0102] Control device FIG. 5 shows a diagram illustrating the flow of control signals and sensor signals between an exemplary control device and an exemplary sample preparation system. In the specific example of FIG. 5, the sample preparation system 1 shown in FIG. 1 is in communication with a control device 50. In FIG. 5, the input stack 11 (see FIG. 1) of the sample preparation system 1 is not shown for ease of understanding the signal flow, but it should be noted that the sample preparation system 1 of FIG. 5 also includes an input stack 11 as shown in FIG. 1.

[0103] As shown in FIG. 5, the control device 50 may include a processor 502 and a storage medium 504 that communicates with the processor 502. The processor 502 may be configured to execute a control process for controlling a sample preparation system such as the exemplary sample preparation system 1. For example, the processor 502 may be configured to communicate with various sensors provided in the sample preparation system 1 to obtain system information from the sensors. Further, for example, the processor 502 may be configured to communicate with components included in the sample preparation system 1 to transmit control signals to the components. The processor 502 may communicate with the sensors and components via one or more wired and / or wireless networks. The processor 502 may further obtain from the storage medium 504 information necessary to execute the control process (e.g., one or more system information other than that which can be obtained from the sensors, one or more values of operation parameters for the control process, etc.).

[0104] As shown in FIG. 5, the control device 50 may receive system information from a first liquid sensor 15, a pump 9 (e.g., a sensor (not shown) provided in the pump 9 to measure the rotational speed of the pump 9), a first pressure sensor 17, a second pressure sensor 18, a second liquid sensor 16, and / or a target substance detection sensor 21. The system information from each sensor may include information indicating the value measured by each sensor.

[0105] The processor 502 can process the system information received from each sensor to control the sample preparation system 1. For example, based on the processing of the system information, the processor 502 determines how at least one component of the sample preparation system 1 should operate, and generates at least one control signal for instructing at least one component to operate as determined, and outputs the generated control signal(s) to at least one component. In some exemplary embodiments, the processor 502 may be configured to function as a PID controller (Proportional-Integral-Derivative controller).

[0106] As shown in FIG. 5, the control device 50 can output control signals to the input sample valve 14, the process fluid input valve 24, the pump 9, the first valve 4, the second valve 5, the output valve 20, and / or the waste pump 25. Additionally, although not shown in FIG. 5, the control device 50 may output control signals to the filter handling device 27 (see FIGS. 2 and 3) and / or the pipetting robot 30 (see FIG. 4).

[0107] The storage medium 504 can be configured to store information necessary for the processor 502 to execute the control process.

[0108] Details of the control process executed by the processor 502 of the control device 50 will be described later.

[0109] Note that FIG. 5 shows the control device 50 having a single processor 502, but in some exemplary embodiments, the control device 50 may have two or more processors 502. Further, FIG. 5 shows the control device 50 as a device separate from the sample preparation system 1, but in some exemplary embodiments, the control device 50 may be incorporated into the sample preparation system 1 as part of the sample preparation system 1.

[0110] Sample Preparation Process As described above, the sample preparation system can perform one or more unit operations such as clarification and purification on a sample.

[0111] In the specific example of FIG. 1, the sample preparation system 1 can perform clarification and / or purification.

[0112] a) Clarification process The clarification process can be understood as a process of clarifying the sample fluid by passing it through a clarification filter (e.g., the first consumable 2a) supported by a first filter holder (e.g., the first means 2) to supply the clarified sample fluid.

[0113] A typical example of the clarification process performed by the sample preparation system 1 is shown below.

[0114] At the beginning of the exemplary clarification process (a), a clarification filter may be provided in the first means 2, and the clarification filter is new, in other words, unused (hereinafter referred to as "step (c1)"). In said step (c1), preferably, the clarification filter is provided by the consumable handling device from the input stack.

[0115] In some cases, the clarification filter may already be connected to the first means 2 at the start of the clarification process (a). In any case, it may be necessary to replace the clarification filter during the clarification process (a) to avoid clogging of the clarification filter and product loss. In some exemplary embodiments, the clarification filter may be automatically discharged from the first means 2 and automatically replaced with a new clarification filter.

[0116] Preferably, based on the measured back pressure of the clarification filter measured by the first pressure sensor 17, or the pressure difference between the first pressure sensor 17 and the second pressure sensor 18, and the pumping speed, it is possible to automatically discharge the clarification filter from the first means and make a decision to automatically replace it with a new clarification filter. Details of how such a decision is made will be described later with reference to the exemplary process shown in FIG. 7.

[0117] Next, the clarification filter is preferably clamped (i.e., "step (c2)") such that the upper and lower surfaces of the clarification filter are supported against the internal pressure during filtration.

[0118] Furthermore, an air pressure test can be performed on the sample preparation system 1 and the clarification filter before the sample to be clarified is pumped into the clarification filter (i.e., "step (c3)"). In particular, in said step (c3), the first valve and the second valve are set to close the flow path, and the pump 9 pumps air into the sample preparation system so as to reach a pre-set pressure. In some situations, the pre-set pressure is from 0.1 bar to 4 bar, preferably from 0.5 bar to 1 bar. The pressure is then monitored for several seconds. At the end of the air pressure test, the pressure is released by setting the first valve, the second valve, and the output valve so that the pressure is released to the waste container via the third line.

[0119] After the optional air pressure test, the sample to be clarified can be supplied to the input cup 13 of the sample preparation system 1 (i.e., "step (c4)a"). Alternatively, the sample to be clarified can also be supplied to the input cup 13 of the sample preparation system 1 before any of steps (c1) to (c3) (i.e., "step (c4)b"). Preferably, the pipetting robot 30 can supply the sample to be clarified to the input cup 13 by transferring the sample to be clarified from at least one input container to the input cup 13 of the sample preparation system 1.

[0120] Next, the sample to be clarified can be pumped from the input cup 13 to the clarification filter (i.e., "step (c5)"). In some situations, in the said step (c5), the sample to be clarified passes through at least the input sample valve 14, the first liquid sensor 15, and the first pressure sensor 17 on the way from the input cup 13 to the clarification filter.

[0121] Furthermore, the sample is clarified by passing it through a clarification filter connected to the first means 2 (hereinafter referred to as "step (c6)").

[0122] Next, the clarified sample can be guided to the output cup 23 (hereinafter referred to as "step (c7)"). In some situations (for example, when the clarified sample does not need to be purified), in the said step (c7), the clarified sample passes through at least the second pressure sensor 18, the second liquid sensor 16, the third line 8, the target substance detection sensor 19, and the output valve 20 from the downstream side of the clarification filter to the output cup 23. In other situations (for example, when the clarified sample needs to be purified), in the said step (c7), the clarified sample passes through at least the second pressure sensor 18, the second liquid sensor 16, the second line 3, the second consumable 3a, the target substance detection sensor 19, and the output valve 20.

[0123] When the input cup 13 is empty, the first liquid sensor 15 can detect that there is no more sample to be clarified (i.e., "step (c8)"). Specifically, the first liquid sensor can detect whether the sample to be clarified previously supplied to the input cup 13 has already passed the position where the first liquid sensor 15 is disposed.

[0124] After step (c8), a rinse step can be automatically executed (i.e., "step (c9)"). In particular, a rinse liquid such as PBS can be supplied to the sample preparation system 1 by one of the process fluid input valves 24 disposed upstream of the first means 2. Thereby, the clarified sample still present upstream of the first means 2 can be induced to the first means 2, and the sample in the clarification filter can be rinsed through the clarification filter. Preferably, the rinse liquid that has passed through the clarification filter and contains the clarified sample is induced to the output cup 23.

[0125] After step (c8) and / or step (c9), preferably, a step of pumping air is executed (i.e., "step (c10)"). Specifically, air can be supplied to the sample preparation system 1 by one of the process fluid input valves 24 disposed upstream of the first means 2. The air pushes the clarified sample or rinse liquid still present in the upper flow field of the clarification filter and the already partially clarified sample or rinse liquid still present in the clarification filter, and passes through the clarification filter. Thereby, the generated and clarified sample can be induced to the output cup 23.

[0126] By applying step (c10), the product recovery rate can be increased. The air pressure used may be in the range of, for example, 0.5 bar to 2 bar. Thereby, the air pressure may be below the bubble point of the clarification filter so as to avoid the formation of bubbles. Typically, the bubble point of the clarification filter is higher than 3 bar. For example, after a waiting time of 0 seconds to 5 seconds, the air pressure is released by reversing the pump 9. The measurement of the bubble point is well known to those skilled in the art. For example, the bubble point can be measured in accordance with ISO 2942 or ASTM F316-03.

[0127] Next, the clarification filter can be discharged from the first means 2 (i.e., "step (c11)"). When the first means 2 is the first filter holder and the clarification filter is clamped by the first clamp 10a and the second clamp 10b, the clarification filter is unclamped before it is discharged. The discharged clarification filter can be received by the filter handling device 27 and supplied to the waste bin 26 (see, for example, FIG. 2).

[0128] Furthermore, in some situations, the connection means (e.g., the hole 29 shown in FIG. 3) included in the filter handling device 27 can connect the input line 12 and the first line 6 to provide fluid communication between the input line 12 and the first line 6 so that the liquid path is closed (i.e., "step (c12)"). This can be achieved, for example, by using an index wheel as the filter handling device and rotating the index wheel to a second position where a through-fluid passage that can then be connected to the input line 12 and the first line 6 is arranged.

[0129] Thereafter, the clarified sample and / or rinse liquid containing the target substance, which is present downstream of the first means, can be induced into the output cup 23 by pumping air (i.e., "step (c13)").

[0130] In some situations, the output sample present in the output cup can be transferred to at least one output container (i.e., "step (c14)"). This step can be performed, for example, by a pipetting robot 30.

[0131] Optionally, the sample preparation system 1 can be cleaned by pumping a cleaning fluid such as a buffer, sodium hydroxide, and / or water from the process fluid input valve 24 disposed upstream of the first means 2 through the sample preparation system 1 to the output cup 23 and then, conversely, from the output cup 23 to the input cup 13 (i.e., "step (c15)"). The resulting liquid can then be directed to a waste container using the waste pump 25.

[0132] b) Purification process The purification process can be understood as a process of purifying the sample fluid by passing it through a purification filter or a purification chromatography column (e.g., the second consumable 3a) connected to the second means 3 to provide a purified sample fluid.

[0133] In some exemplary embodiments, the purification process may be performed after the clarification process as described above. However, in some situations, the purification process may be performed without performing the clarification process, in which case the clarification filter is not set in the filter holder (e.g., the first means 2), and the fluid may flow from the input line 12 to the first line 6 through at least the holes 29 of the filter handling device 27 (see FIGS. 1 and 3) without passing through the clarification filter.

[0134] Control process The clarification process as described above can be driven at a pump flow rate having a maximum differential pressure (which can also be referred to as "dP") across the clarification filter (e.g., the first consumable 2a) to maintain the integrity of the cells. When the maximum differential pressure is reached, the control device 50 can control the pump 9 to decelerate to keep the differential pressure constant. When the clarification process is controlled by the differential pressure by reducing the flow rate (e.g., by reducing the rotational speed of the pump 9), the clarification process can be very slow.

[0135] When the maximum differential pressure is reached, the clarification process may have a diminishing return and a process stop point, such as a minimum flow rate or a maximum process time, must be determined. It is desirable to minimize the DNA and host cell protein (HCP) in the output sample. This can be achieved by maintaining the differential pressure as low as possible.

[0136] For maximum recovery, a rinse of the clarification may be required. The ideal balance for maximum efficiency is to time the switch from the input culture medium to the rinse buffer such that the maximum differential pressure is just reached when the rinse is complete, but the control device 50 does not need to reduce the pump speed (e.g., the rotational speed of pump 9). In this scenario, the clarification process may operate at maximum speed for as long as possible. This can be technically difficult as the fouling of the clarification filter is non-linear and may depend on the "filterability" of the input culture medium.

[0137] The control process executed by the control device 50 in accordance with the present disclosure can achieve or (at least come close to achieving) such clarification optimal conditions. The control process executed by the control device 50 in accordance with the present disclosure can also avoid the worst-case scenario of filter blockage, which may reduce the product recovery rate over time and may require further intervention to pass through.

[0138] FIG. 6 shows a graph of exemplary recorded online data obtained by running an exemplary sample preparation system 1 without the control process according to the present disclosure. FIG. 6 shows an example of the effect of waiting until the maximum differential pressure limit of 1000 mbar. In other words, in the example of FIG. 6, the control device 50 controls pump 9 of the sample preparation system 1 to maintain the sample fluid flow into the flow path towards the clarification filter while the differential pressure is below the 1000 mbar limit and reduce the rotational speed of pump 9 (e.g., the pump flow rate) when the differential pressure reaches the 1000 mbar limit.

[0139] As shown in Figure 6, the pump flow rate (shown as "Liquid Controller Pump Instantaneous Speed" in Figure 6) decreases, the process timing becomes slower, and it hinders the rinse buffer from easily passing through the filter (see, for example, "Total Volume Pumped Through Valve (Equilibrium / Rinse 1)" in Figure 6). It is impossible to completely rinse the filter, and thus the product is retained in the filter, which can thereby reduce the product recovery rate. Further, this can increase cell lysis (which can be indicated by LDH (lactate dehydrogenase)) and HCP contaminants in the output.

[0140] Figure 7 shows a flowchart of an exemplary process for controlling a sample preparation system. More specifically, the exemplary process shown in Figure 7 can be executed to control the sample preparation system 1 when the sample preparation system 1 performs a clarification process as described above.

[0141] The exemplary process shown in Figure 7 can be executed by the processor 502 of the control device 50. The exemplary process shown in Figure 7 can be started when instructions for starting the exemplary process are received from a user via an input device (not shown). The instructions from the user can include, for example, instructions for the sample preparation system to start clarifying a sample fluid.

[0142] As will be apparent from the description of the exemplary process shown in Figure 7, it should be noted that the clarification process executed in the sample preparation system 1 when the processor 502 controls the sample preparation system 1 according to the exemplary process shown in Figure 7 is not exactly the same as a typical example of the above-described clarification process. Specifically, for example, the rinse step (c9) and / or the pneumatic pumping step (c10) may be executed under different conditions from the step (step (c8)) of detecting that there is no more sample to be clarified.

[0143] When the exemplary process shown in FIG. 7 starts, at step S10, the processor 502 may receive one or more operating parameters for the exemplary process. The one or more operating parameters may include at least a specified maximum differential pressure. The specified maximum differential pressure may be input by the user using an input device (not shown) connected to or communicating with the processor 502.

[0144] In some exemplary embodiments, the one or more operating parameters may further include a specified period Δt (represented by a unit of time (e.g., seconds, minutes, etc.) or the amount of fluid introduced into the fluid path (e.g., volume)) used when determining a regression function, as will be described in detail below with respect to step S16. In some exemplary embodiments, the one or more operating parameters may further include a specified stop condition (e.g., a first specified condition) for determining whether to stop introducing the sample fluid into the fluid path of the sample preparation system 1, as will be described later with respect to step S18.

[0145] The specified period Δt and / or the specified stop condition may also be input by the user using an input device.

[0146] Optionally, the one or more operating parameters may further include one or more of the cell density of the sample fluid, the turbidity of the sample fluid, the particle size distribution of the sample fluid, the optical density of the sample fluid, the filter resistance index, and the filter surface area. The values of these additional operating parameters may be input by the user using an input device. After step S10, the exemplary process can proceed to step S12.

[0147] In step S12, the processor 502 may set the valve positions and pump flow rates to initiate the clarification process. For example, the processor 502 may generate a control signal to set the process fluid input valve 24 so as to prevent various types of process fluids from flowing into the fluid path, and output the generated control signal to each process fluid input valve 24. Further, the processor 502 may generate a control signal to set the input sample valve 14 so that the sample fluid flows from the input cup 13 towards the pump 9 and the first means 2, and output the generated control signal to the input sample valve 14. Further, the processor 502 may generate a control signal to instruct the pump 9 to start pumping the fluid at a specified rotational speed (for example, a default rotational speed according to the type of sample fluid, a rotational speed specified by the user, etc.), and output the generated control signal to the pump 9.

[0148] Further, for example, the processor 502 may generate a control signal to set the first valve 4, the second valve 5, and / or the output valve 20 to a position suitable for the desired flow of the clarified sample fluid, and output the generated control signal to the first valve 4, the second valve 5, and / or the output valve 20, respectively. For example, when the clarified sample fluid undergoes a purification process, the first valve 4 is set to allow the clarified sample fluid to flow towards the second means 3.

[0149] Further, for example, when the clarified sample fluid is collected in the output cup 23 without a purification process, the first valve 4, the second valve 5, and the output valve 20 are set to allow the clarified sample fluid to flow towards the output cup 23 through the third line 8 and the output line 22. In summary, the processor 502 may control the sample preparation system 1 to perform steps (c5), (c6), and (c7) of the exemplary clarification process as described above. After step S12, the exemplary process may proceed to step S14.

[0150] In step S14, the processor 502 may monitor sensors provided on the sample preparation system 1 (for example, while the sample preparation system 1 executes steps (c5), (c6), and (c7) of the exemplary clarification process described above). For example, the processor 502 may receive system information indicating at least the amount of sample fluid introduced into a fluid path (for example, including at least the input line 12 and the first line 6), the upstream pressure at the first pressure sensor 17, and the downstream pressure at the second pressure sensor 18.

[0151] The amount of sample fluid introduced into the fluid path may be obtained, for example, from a sensor (not shown) on the pump 9 for measuring the rotational speed of the pump 9. For example, the amount of sample fluid may be calculated using the rotational speed of the pump 9 and the time elapsed since the start of the sample preparation process executed by the sample preparation system 1. More specifically, for example, when the rotational speed of the pump 9 is represented by the amount of fluid pumped by the pump 9 per unit time (for example, ml / min), the amount of sample fluid introduced into the fluid path may be the product of the average rotational speed of the pump 9 and the time elapsed since the start of the sample preparation process. Further, in some embodiments, the amount of sample fluid introduced into the fluid path may be proportional to the number of rotations of the pump 9 since the start of the sample preparation process (for example, the number of revolutions). Therefore, if the amount of fluid that can be pumped by one rotation of the pump 9 is known, the amount of sample fluid can be obtained by calculating the number of rotations of the pump 9 since the start of the sample preparation process from the rotational speed of the pump 9 (for example, represented by rpm) and the elapsed time.

[0152] In some exemplary embodiments, the rotational speed sensor may not necessarily be provided on the pump 9. In such exemplary embodiments, a calibrated value of the rotation of the pump 9 (driven, for example, by a DC motor or a stepper motor) may be obtained from the storage medium 504, and the amount of sample fluid may be calculated using the obtained calibrated value of the rotation of the pump 9.

[0153] Further, in some exemplary embodiments, instead of obtaining the downstream pressure from the second pressure sensor 18, the atmospheric pressure stored in the storage medium 504 may be obtained as the downstream pressure.

[0154] During the monitoring in step S14, the processor 502 may repeatedly receive system information (specifically, one or more values that can change over time, such as the amount of fluid introduced into the fluid path, the upstream pressure, etc.) at a specified time interval (e.g., 500 ms, but it may be longer or shorter depending on the processing capacity). If the system information includes one or more values that do not change over time (such as the atmospheric pressure as the downstream pressure), it may not be necessary for such values to be repeatedly received by the processor 502, and it may be sufficient to receive them at least once. After step S14, the exemplary process can proceed to step S16.

[0155] In step S16, the processor 502 may determine a regression function for extrapolating when the clarification filter will become blocked. For example, the regression function is based on the differential pressure dP_t across the clarification filter obtained for different time points t i -Δt,…,t i with respect to the upstream pressure and the downstream pressure within the specified period Δt, and the differential pressure dP_t i -Δt,…,dP_t i is the value of the dependent variable y of the regression function, and the amount of sample fluid Vin_t introduced into the fluid path obtained for different time points t i -Δt,…,t i within the specified period Δt is assumed to be the value of the independent variable x of the regression function, and thus may be determined. Therefore, the regression function can be determined using the system information obtained (in step S14) for the specified period Δt preceding the current time point t. As also described above, the current time point t i -Δt,…,Vin_t i can be represented by the amount of fluid introduced into the fluid path so far since the start of the sample preparation process executed by the sample preparation system 1. Alternatively, the current time point t current can be represented by the amount of fluid introduced into the fluid path so far since the start of the sample preparation process executed by the sample preparation system 1. Alternatively, the current time point t current can be represented by the amount of fluid introduced into the fluid path so far since the start of the sample preparation process executed by the sample preparation system 1. Alternatively, the current time point t currentmay be represented in units of time (e.g., t seconds or minutes from the start of the sample preparation process).

[0156] The specified period Δt used in step S16 may vary according to the sample preparation process executed by the sample preparation system 1. In some situations, the specified period Δt may be as short as the monitoring interval (step S14), e.g., 500 ms. In other situations, the specified period Δt may be the total duration of the sample preparation process, in other words, the length from the start of sample preparation to the current time. In some exemplary embodiments, the specified period Δt may be the period during which 1.0 ml of fluid is introduced into the fluid path, or 10 seconds.

[0157] In some exemplary embodiments, the specified period Δt may be immediately before the current time t current Here, "immediately before" may indicate the period up to the time point of a single time interval before the current time (for receiving system information). For example, if the time interval for receiving system information during the monitoring step S14 is 500 ms and the specified period is 10 seconds, the specified period Δt immediately before the current time t current may be the period starting from t current - 10.5 seconds and ending at t current - 0.5 seconds.

[0158] In some other exemplary embodiments, the specified period Δt may not necessarily be immediately before the current time t current For example, the specified period Δt may be before the current time t current and preferably indicate the period up to a point within a specific ratio (e.g., 1 / 10) of the expected total time of the sample preparation process (or the total amount of sample fluid).

[0159] In some exemplary embodiments, the regression function may be a linear regression function as follows. y = ax + c (1), Here, x and y may be an independent variable and a dependent variable as described above, respectively, and a and c may be coefficients of the regression function. From the monitoring of the system information in step S14, pairs of x-y values during a specified period can be obtained.

[0160] More specifically, a pair can be obtained between the amount of sample fluid introduced into the fluid path during the specified period (e.g., the value of x) and the differential pressure across the clarification filter (e.g., the value of y that can be determined by calculating the difference between the downstream pressure of the second pressure sensor 18 and the upstream pressure of the first pressure sensor 17). The coefficients a and b of the regression function (1) can be determined using the pairs of x-y values obtained during the specified period.

[0161] In some further exemplary embodiments, the regression function may have a form different from the linear regression function as described above. For example, one of the following regression functions may be used as the regression function in step S16. y = ax b + c (2), y = ae bx + c (3), Here, x and y may be an independent variable and a dependent variable as described above, respectively, and a, b, and c may be coefficients of the regression function. Also, when function (2) or function (3) is used as the regression function in step S16, the coefficients a, b, and c may be determined using the pairs of x-y values obtained during the specified period as described above.

[0162] In an exemplary embodiment in which one or more additional operating parameters (e.g., one or more of cell density, turbidity, particle size distribution, optical density, filter resistance index, filter surface area) are obtained, the additional operating parameters can also be used when determining the regression function.

[0163] For example, one or more additional operation parameters may be used to determine one or more values of coefficients included in a regression function, such as coefficients a, b, and / or c of the above-described regression functions (1), (2), and (3). Further, for example, one or more additional operation parameters may be used to determine or adjust a specified period during which system information is used to determine a regression function.

[0164] More specifically, for example, for each of one or more additional operation parameters, the processor 502 determines, as a function of one of the one or more parameters, a blocking amount formula for determining a blocking amount, and a blocking amount indicating an amount of sample fluid that can block the flow of the sample fluid to at least one consumable when introduced into at least one consumable.

[0165] Further, for example, for each of one or more additional operation parameters, the processor 502 the amount of sample fluid currently introduced into the fluid path, or the total amount of sample fluid introduced into the fluid path, and a coefficient indicating a ratio between the predicted blocking amount obtained using the generated formula and may be determined.

[0166] Subsequently, the processor 502 may determine a regression function using the coefficient determined for each of one or more additional operation parameters.

[0167] To determine a regression function, for example, the processor 502 determines a sensitivity coefficient using the coefficient determined for each of one or more additional operation parameters, and based on the sensitivity coefficient, selects one type of regression function to be used as the regression function from a plurality of types of regression functions.

[0168] In a specific example, the sensitivity coefficient can be calculated as follows.

Equation

[0169] Coefficient α i When it is equal to 1, the volume V of the sample fluid input may be equal to the predicted occlusion volume V predicted_i When the coefficient α i is less than 1, the volume V of the sample fluid input may be less than the predicted occlusion volume V predicted_i When the coefficient α i is greater than 1, the volume V of the sample fluid input may be greater than the predicted occlusion volume V predicted_i The number N of additional operation parameters to be used may be specified by the user, for example, using an input device (not shown) that communicates with the processor 502.

[0170] In some exemplary embodiments, when the sensitivity coefficient determined as described above is greater than a specified sensitivity threshold, the processor 502 may select a more sensitive (in other words, more reactive) regression function from a plurality of types of regression functions. On the other hand, when the sensitivity coefficient is less than or equal to the specified sensitivity threshold, the processor 502 may select a less sensitive (in other words, less reactive) regression function from a plurality of types of regression functions.

[0171]

[0172] ​For example, when multiple types of regression functions include the regression functions (1), (2), and (3) as described above, the linear regression function (1) may be the least sensitive among the three. Which of the regression functions (2) and (3) is more sensitive may depend on the values of the coefficients a, b, and c. So, if a more sensitive regression function is selected (for example, the sensitivity coefficient is greater than the specified sensitivity threshold), one or more of the coefficients a, b, and c may be set to different values, or a more sensitive regression function with a specific value (one or more) of the coefficients a, b, and / or c may be selected. The specified sensitivity threshold can be determined in advance using test data obtained from experiments.

[0173] Furthermore, the sensitivity coefficient may be used to determine and / or adjust the specified period during which system information is used to determine the regression function. For example, the specified period may be set longer for a lower sensitivity coefficient than for a higher sensitivity coefficient.

[0174] After step S16, the exemplary process can proceed to step S18.

[0175] In step S18, the processor 502 may determine whether the specified stop condition is satisfied. For example, when the value indicating the predicted blockage volume Vpb is current the amount of sample fluid Vin_t introduced into the fluid path at the current time t current it may be determined that the specified stop condition is satisfied. The predicted blockage volume Vpb may be the value of the independent variable x of the regression function when the value of the dependent variable y is the specified maximum differential pressure.

[0176] The predicted blockage volume Vpb can be understood as the predicted amount of sample fluid introduced into the filter when the differential pressure across the filter reaches the specified maximum differential pressure. The predicted blockage volume may also be referred to as the predicted filter blockage volume, or, when the amount is represented by volume, the predicted filter blockage volume. Furthermore, the amount of sample fluid Vin_t introduced into the fluid path at the current time t current currentIt can also be referred to as the current sample amount, or when the amount is represented by volume, the current sample volume.

[0177] In some exemplary embodiments, the processor 502 may determine that the specified stop condition is satisfied when the difference between the predicted filter clogging amount and the current sample amount (e.g., (predicted filter clogging amount) - (current sample amount)) is less than or equal to a specified threshold value (e.g., 2 ml). The processor 502 may determine that the specified stop condition is not satisfied when the difference between the predicted filter clogging amount and the current sample amount is greater than the specified threshold value. The specified threshold value may be related to the input volume of the sample preparation system, which may be determined by the volume of the (one or more) conduits between the input valve 14 and the clarification filter (e.g., filter 2a).

[0178] The specified threshold value may also be received by the processor 502 in step S10. In exemplary embodiments where (one or more) additional operating parameters are used as described above, the specified threshold value may be adjusted using the additional operating parameters. Alternatively or additionally, the specified threshold value may be adjusted using system information obtained with respect to the specified period Δt used to determine the regression function in step S16. For example, if the predicted filter clogging volume Vpb obtained from the regression function decreases as the repetition of step S16 progresses, the specified threshold value may be increased. Further, if the predicted filter clogging volume Vpb obtained from the regression function increases as the repetition of step S16 progresses, the specified threshold value may be decreased. Here, it should be noted that the predicted filter clogging volume Vpb obtained from the regression function may depend on system information indicating the upstream pressure and the downstream pressure.

[0179] If the processor 502 determines that the specified stop condition is not satisfied (No in step S18), the exemplary process of FIG. 7 can return to step S14. In this case, the monitoring step S14, the determination of the regression function in step S16, and the determination of whether the specified stop condition is satisfied in step S18 are repeated, during which the processor 502 controls the sample preparation system 1 to continue introducing the sample fluid into the fluid path of the sample preparation system 1. Since the system information received for different periods can be used to determine the regression function, it should be noted that the regression function determined in step S16 can be different from that determined in different iterations of steps S14, S16, and S18. Specifically, since the specified period Δt used in step S16 can shift forward as time elapses, the system information used in step S16 can include different information in different iterations. Therefore, the regression function can be updated in real time.

[0180] If the processor 502 determines that the specified stop condition is satisfied (Yes in step S18), the exemplary process can proceed to step S20.

[0181] In step S20, the processor 502 can control the sample preparation system 1 to stop introducing the sample fluid into the fluid path of the sample preparation system 1. For example, the processor 502 can generate a control signal for setting the input sample valve 14 to a closed state in which the sample fluid is prevented from flowing toward the filter in the input line 12, and output the generated control signal to the input sample valve 14. Further, for example, the processor 502 can generate a control signal for instructing the pump 9 to stop moving the sample fluid in the input line 12, and output the generated control signal to the pump 9. After step S20, the exemplary process can proceed to step S22 or step S24.

[0182] In optional step S22, the processor 502 may control the sample preparation system 1 to pump the rinse liquid through the filter (e.g., the rinse step (c9) of the exemplary clarification process as described above). For example, the processor 502 may generate a control signal to set one of the process fluid input valves 24 corresponding to the source of the rinse liquid to an open state that allows the rinse liquid to flow toward the filter within the fluid path, and output the generated control signal to one of the process fluid input valves 24.

[0183] Further, for example, the processor 502 may generate a control signal to instruct the pump 9 to move the fluid (e.g., the rinse liquid and any remaining sample fluid) within the input line 12 toward the filter, and output the generated control signal to the pump 9. The sample fluid remaining in the input line 12 and / or the filter by step S22 may pass through the filter together with the rinse liquid. Further, the processor 502 may set one of the process fluid input valves 24 to a closed state that prevents the rinse liquid from flowing into the input line 12 at the end of the rinse step S22 (e.g., after a specific period). After step S22, the exemplary process may proceed to step S24.

[0184] Note that step S22 is an optional step and may be skipped in some exemplary embodiments.

[0185] In step S24, the processor 502 may control the sample preparation system 1 to flush air through the filter (e.g., the air pumping step (c10) of the exemplary clarification process as described above). For example, the processor 502 may generate a control signal to set one of the process fluid input valves 24 corresponding to the source of air to an open state that allows air to flow toward the filter within the fluid path (e.g., the input line 12), and output the generated control signal to one of the process fluid input valves 24.

[0186] Furthermore, for example, the processor 502 may generate a control signal for instructing the pump 9 to move the fluid (e.g., air as well as the rinse liquid and / or any remaining sample fluid) in the input line 12 towards the filter, and output the generated control signal to the pump 9. The sample fluid and / or the rinse liquid remaining in the input line 12 and / or in the filter by step S24 may pass through the filter together with the air. Furthermore, the processor 502 may set one of the process fluid input valves 24 to a closed state in which air is prevented from flowing into the input line 12 at the end of the air flush step S22 (e.g., after a specific period). After step S24, the exemplary process may proceed to step S26.

[0187] In step S26, the processor 502 may control the sample preparation system 1 to discharge the filter (e.g., step (c11) of the exemplary clarification process as described above). For example, the processor 502 may control the filter handling device 27 (see, e.g., FIGS. 2 and 3) to remove the filter from the first means 2. More specifically, the processor 502 may generate a control signal for removing the filter from the first means 2 by receiving the filter, and output it to the filter handling device 27. Thus, the filter can be automatically removed from the fluid path of the sample preparation system 1. After step S26, the exemplary process may proceed to step S28.

[0188] In step S28, the processor 502 may determine whether to set another filter (e.g., a new, unused filter) in the fluid path of the first means 2. For example, the processor 502 may determine to set another filter when a specified replacement condition (e.g., a second specified condition) is satisfied, or the processor 502 may determine not to set another filter when the specified replacement condition is not satisfied. The specified replacement condition may include a condition related to the amount of remaining sample fluid to be processed by the sample preparation system 1. For example, a threshold value of the amount of remaining sample fluid may be specified (e.g., by the user), and the specified replacement condition is determined to be satisfied when the amount of remaining sample fluid exceeds the threshold value.

[0189] If the processor 502 determines to set another filter (Yes in step S28), the exemplary process can proceed to step S30.

[0190] In step S30, the processor 502 may control the sample preparation system 1 to set a new filter (e.g., an unused filter) in the fluid path of the first means 2. For example, the processor 502 may generate a control signal for setting a new filter in the first means 2 and output it to the filter handling device 27. After step S30, the exemplary process can return to step S12.

[0191] If the processor 502 determines not to set another filter (No in step S28), the exemplary process may end.

[0192] After or in parallel with the exemplary process shown in FIG. 7, the processor 502 may perform a purification process using the second means 3a (e.g., a purification filter or a purification chromatography column), direct the fluid from the first means 2 or the second means 3 towards the output cup 23 or the waste container, and execute one or more further control processes to control the sample preparation system 1 to, for example, clean the sample preparation system 1.

[0193] Experimental results FIG. 8 shows a graph of exemplary recorded online data obtained by running an exemplary sample preparation system controlled by the control process according to the present disclosure. In the example shown in FIG. 8, the control device 50 executed the exemplary process shown in FIG. 7 using the above-described linear regression function (1) with a specified period of 10 seconds or a change in the volume of the fluid introduced into the fluid path of 2 ml. As can be seen from FIG. 8, the filter stops treating the culture fluid at 650 mbar so as not to reach the limit of 1000 mbar before flowing the entire rinse fluid. The differential pressure ("Calculated_dp_Clarification_Pressure" in FIG. 8) increases up to 800 mbar during rinsing, but does not clog the filter, and the flow rate ("Liquid Controller Pump Instantaneous Speed" in FIG. 8) remains at the set point.

[0194] FIG. 9 shows a graph showing the clogging volume of the filter provided on the fluid path of the sample preparation system predicted by the control process according to the present disclosure, as well as the volume pumped through the input sample valve 14 and the differential pressure across the filter. The control device 50 executed the exemplary process of FIG. 7 using the above-described linear regression function (1) with a specified period of 10 seconds or a change in the volume of the fluid introduced into the fluid path of 2 ml. The maximum differential pressure was 1000 mbar. In this example of FIG. 9, the specified threshold was not met by the difference between the predicted filter clogging volume and the current sample volume. In other words, the sample fluid ran out and no filter clogging occurred.

[0195] Figure 10 shows a further exemplary graph showing the blocked volume of a filter provided on the fluid path of a sample preparation system predicted by the control process according to the present disclosure, as well as the volume pumped through the input sample valve 14 and the differential pressure across the filter. The control device 50 executed the exemplary process of FIG. 7 using the linear regression function (1) as described above. The maximum differential pressure was set to 1000 mbar. As can be seen from FIG. 10, the difference between the actual input sample volume (shown as "volume pumped through valve - input") and the predicted filter blocked volume (shown as Predicted_Blockage_Volume) converges towards a smaller value than at the start of the process. Thus, this difference satisfied a specified threshold, and the sample preparation system was controlled to stop the flow of sample fluid into the fluid path.

[0196] Figure 10 shows two exemplary periods t that can be used to determine the regression function in two different iterations of steps S14, S16, and S18 of the exemplary process of FIG. 7 i from -Δt to t i and t j from -Δt to t j are further shown. For example, the differential pressure dP_t i acquired for the period from -Δt to t i -Δt…, dP_t i and the input volume Vin_t i -Δt,…, Vin_t i -Δt,…, Vin_t i can be used to determine the regression function at time point t i (or a time point after time point t i ). Further, the regression function determined at time point t i (or a time point after time point t i ) can be used to calculate the predicted blocked volume Vpb_t i at time point t i (or the predicted blocked volume Vpb at a time point after time point t i ). Similarly, for example, the differential pressure dP_t j acquired for the period from -Δt to t j -Δt,…, dP_t j -Δt,…, dP_tj and the input volume Vin_t j -Δt, …, Vin_t j is at time t j (or at a time after time t j may be used to determine a regression function at time t (or at a time after time t). Further, the regression function determined at time t j (or at a time after time t j may be used to calculate the predicted occlusion volume Vpb_t at time t j (or the predicted occlusion volume Vpb at a time after time t). j (or at a time after time t j (or the predicted occlusion volume Vpb at a time after time t).

[0197] In the specific example of FIG. 10, the specified stop condition (see step S18 in FIG. 7) was satisfied at time t j . For this reason, when the sample preparation system 1 is controlled to stop introducing the sample fluid into the fluid path, the differential pressure dP after time t j decreased. Further, when the pump 9 is controlled to release the differential pressure, the input sample volume Vin after time t j also slightly decreased.

[0198] Modification In the above exemplary embodiment, the sample preparation system 1 as shown in FIG. 1 is controlled by the control device 50.

[0199] In a further exemplary embodiment, a sample preparation system having a configuration different from the example shown in FIG. 1 may be controlled by the control device 50. For example, the sample preparation system to be controlled may include a unit configured to perform clarification instead of a unit configured to perform purification.

[0200] Furthermore, in some exemplary embodiments, the controlled sample preparation system may be configured to perform filtration operations other than the clarification process. In such cases, the filter provided on the fluid path is not a clarification filter but a filter for different filtration operations. For example, the sample preparation system may perform any biological filtration process of biomolecules using filters such as syringe filters, ultrafiltration filters, membrane filters, glass and quartz microfiber filters, syringe-less filters, filter papers, hollow fiber filters, etc. Further, for example, when a membrane filter is used, the filter may have different molecular weight cut-offs (MWCO) such as 1 kDa, 5 kDa, 10 kDa, 30 kDa, 50 kDa, 100 kDa, 300 kDa, 500 kDa, 0.2 μM, etc., depending on the purpose of the filtration operation.

[0201] Furthermore, in some exemplary embodiments, the controlled sample preparation system may comprise two or more series arrangements of the first means 2 and the second means 3, in other words, two or more series arrangements of units for different unit operations (e.g., clarification and purification). In particular, each of these series arrangements may have the same configuration as the series arrangement described above with respect to the sample preparation system 1 shown in FIG. 1. If there are two or more series arrangements within the sample preparation system, the series arrangements may be arranged in parallel with each other, and each series arrangement may have its own input cup and output cup to enable independent operation. Further, the control device 50 may execute the exemplary processes shown in FIG. 7 in parallel for each of the series arrangements.

[0202] Furthermore, in the above exemplary embodiments, the sample preparation system to be controlled includes an input sample valve that permits or prevents the sample fluid from flowing into the fluid path of the sample preparation system. In further exemplary embodiments, permitting or preventing the sample fluid from flowing into the fluid path of the sample preparation system can be achieved only by controlling the pumps on the fluid path without a valve. In such further exemplary embodiments, a dedicated pump may be provided for each of the input fluid lines (e.g., for the sample fluid and for different types of process fluids) without a valve. The input fluid lines may be connected to each other downstream of the pumps to form a single downstream fluid line, and a liquid sensor may be provided on the single downstream line. The fluid pumped into the fluid path of the sample preparation system can be selected by operating the corresponding pump while stopping other pumps.

[0203] Furthermore, in the above exemplary embodiments, the regression functions (1), (2), and / or (3) may be used. In further exemplary embodiments, an improved prediction model for predicting the filter clogging amount may be used. For example, the prediction model can be improved by identifying replicate samples such that the prediction model is improved for each sample and enabling machine learning (e.g., neural networks, fuzzy logic). The model can also have the potential to improve its prediction ability by incorporating complex filter clogging formulas (e.g., solid adsorption models, etc.).

[0204] Furthermore, the prediction model can take into account all the pipe lengths and dead volumes (including those of the clarification filters) of the sample preparation system, which enables the implementation of the prediction model in different systems and different filters.

[0205] Furthermore, in some exemplary embodiments, prediction filter clogging data may be generated across multiple platforms, filters, and scales, thereby enabling a fully automated scaling software platform for filtration.

[0206] Hardware Configuration FIG. 11 shows an exemplary hardware configuration of a computer that can be used to implement at least a part of the system as described above. For example, at least a part of the control device 50 shown in FIG. 5 can be implemented using the computer 7 shown in FIG. 11. The computer 7 shown in FIG. 11 includes a central processing unit (CPU) 70, a system memory 72, a network interface 74, a hard disk drive (HDD) interface 76, an external disk drive interface 78, and an input / output (I / O) interface 80. These components of the computer are coupled to each other via a system bus 82. The CPU 70 can execute arithmetic operations, logical operations, and / or control operations by accessing the system memory 72. The system memory 72 can store information and / or instructions for use in combination with the CPU 70. The system memory 72 can include volatile and non-volatile memories such as a random access memory (RAM) 720 and a read-only memory (ROM) 722. A basic input / output system (BIOS) including routines useful for transferring information between elements within the computer 7 during startup and the like can be stored in the ROM 722. The system bus 82 can be any of several types of bus structures including a memory bus or memory controller, a peripheral bus, and a local bus using any of a variety of bus architectures.

[0207] The computer can include a network interface 74 for communicating with other computers and / or devices via a network.

[0208] Furthermore, the computer can include a hard disk drive (HDD) 84 for reading from and writing to a hard disk (not shown), and an external disk drive 86 for reading from and writing to a removable disk (not shown). The removable disk may be a magnetic disk for a magnetic disk drive, or an optical disk such as a CD-ROM for an optical disk drive. The HDD 84 and the external disk drive 86 are each connected to the system bus 82 by an HDD interface 76 and an external disk drive interface 78, respectively. The drives and their associated computer-readable media provide non-volatile storage for computer-readable instructions, data structures, program modules, and other data for a general-purpose computer. The data structures can include related data for implementing the exemplary methods described herein and their variations. The related data may be organized in a database, such as a relational or object database.

[0209] Although the exemplary environment described herein uses hard disks (not shown) and external disks (not shown), those skilled in the art should understand that other types of computer-readable media, such as magnetic cassettes, flash memory cards, digital video disks, random access memory, read-only memory, etc., which can store data accessible by a computer, can also be used in the exemplary operating environment.

[0210] The hard disk, external disk, ROM 722, or RAM 720 can store several program modules, including an operating system (not shown), one or more application programs 7202, other program modules (not shown), and program data 7204. The application programs can include at least some of the functions described above.

[0211] Computer 7 can be connected to input devices 92 such as a mouse and / or keyboard and display devices 94 such as a liquid crystal display via corresponding I / O interfaces 80a and 80b and system bus 82. When computer 7 is implemented as a tablet computer, for example, a touch panel for displaying information and receiving input may be connected to computer 7 via a corresponding I / O interface and system bus 82. Further, in some examples, although not shown in FIG. 11, computer 7 may be further connected to imaging devices such as a printer and / or camera via corresponding I / O interfaces and system bus 82.

[0212] In addition to, or as an alternative to, implementations using computer 7 as shown in FIG. 11, some or all of the functions of the exemplary embodiments described herein may be implemented as one or more hardware circuits. Examples of such hardware circuits can include, but are not limited to, large scale integration (LSI), reduced instruction set circuits (RISC), application specific integrated circuits (ASIC), and field programmable gate arrays (FPGA).

[0213] Further aspects The following items provide further aspects of the present disclosure.

[0214] Item 1. A sample preparation system, a series arrangement of a first means configured to be connected to a first consumable and a second means configured to be connected to a second consumable, a first valve disposed between the first means and the second means, a second valve disposed downstream of the second means, a first line providing fluid communication between the first means and the first valve, a second line providing fluid communication between the first valve and the second valve via the second means, A third line configured to bypass the second means and provide direct fluid communication between the first valve and the second vent; A pump configured to move fluid within the sample preparation system; A sample preparation system comprising the same.

[0215] Item 2. The first means is connected to the first consumable and is configured to instruct the first consumable. The sample preparation system according to Item 1.

[0216] Item 3. The second means is connected to the second consumable and is configured to instruct the second consumable. The sample preparation system according to Item 1 or 2.

[0217] Item 4. The first consumable is the first filter. The sample preparation system according to any one of Items 1 to 3.

[0218] Item 5. The second consumable is the second filter or a chromatography column. The sample preparation system according to any one of Items 1 to 4.

[0219] Item 6. The first means is the first filter holder. The sample preparation system according to any one of Items 1 to 5.

[0220] Item 7. A consumable handling device connected to the first means configured to provide the first consumable to the first means and receive the first consumable from the first means The sample preparation system according to any one of Items 1 to 6, further comprising the same.

[0221] Item 8. Further comprising an input line providing fluid communication between an input cup disposed upstream of the first means and the first means, The consumable handling device further comprises connection means configured to connect the input line and the first line to provide fluid communication between the input line and the first line. The sample preparation system according to item 7.

[0222] Item 9. Further comprising an input sample valve disposed downstream of the input cup on the input line and upstream of the first means. The sample preparation system according to item 8.

[0223] Item 10. Further comprising a target substance detection sensor disposed downstream of the second means configured to detect the target substance. The sample preparation system according to any one of items 1 to 9.

[0224] Item 11. A method for preparing a sample using the sample preparation system according to any one of items 1 to 10, wherein the method comprises (i) a step of clarifying the sample by passing the sample through a first consumable connected to the first means to supply a clarified sample, wherein the first consumable is a clarification filter, and (ii) a step of purifying the sample by passing the sample through a second consumable connected to the second means to supply a purified sample, wherein the second consumable is a purification filter or a purification chromatography column including at least one of them.

[0225] Item 12. The method includes steps (i) and (ii), step (i) is performed before step (ii), and the clarified sample flows directly from the clarification filter to the purification filter or the purification chromatography column. The method for preparing a sample according to item 11.

[0226] Item 13. The method includes, between step (i) and step (ii), in the following order: (a) a step of discharging the clarification filter, and (b) a step of providing a through-flow path connecting the line upstream of the first means and the first line of the sample preparation system within the first means. Step of removing the sample from the line upstream of the first means by inducing the sample through the through-flow path, the first line, and the third line of the sample preparation system; The method for preparing a sample according to item 12, further comprising.

[0227] Item 14. The sample is a cell culture solution containing protein, The clarification filter is a cell culture solution clarification filter, The purification filter is a protein purification filter or a protein purification chromatography column, The method for preparing a sample according to any one of items 11 to 13.

[0228] Item 15. In step (i), the clarification filter is automatically discharged from the first means and automatically replaced with a new clarification filter. The method for preparing a sample according to any one of items 11 to 14.

Claims

1. A computer-implemented method for controlling a sample preparation system (1) for preparing a chemical, pharmaceutical, and / or biotechnological sample, wherein the sample preparation system (1) comprises a fluid path and at least one consumable (2a), such as a filter, provided on the fluid path, and the method is: A step of controlling the sample preparation system (1) so that a sample fluid containing a target substance flows through the fluid path through the at least one consumable (2a), wherein the sample fluid can pass through the upstream point in the fluid path, the at least one consumable (2a), and the downstream point in the fluid path in this order. at least The amount of the sample fluid induced into the fluid path, The upstream pressure at the aforementioned upstream point, and Downstream pressure at the aforementioned downstream point Step (S14) to obtain system information indicating, At present time (t ｃｕｒｒｅｎｔ A step of determining a regression function using the system information obtained for a specified period (Δt) preceding ), wherein the regression function is Different point in time (t) within the specified period (Δt) ｉ -Δt, ..., t ｉ The differential pressure (dP_t) across the at least one consumable (2a) is obtained based on the upstream and downstream pressures. ｉ -Δt, ..., dP_t ｉ ) is the value of the dependent variable (y) of the regression function, For different time points (t ｉ −Δt, …, t ｉ ) within the specified period (Δt), the amount of the sample fluid (Vin_t ｉ −Δt, …, Vin_t ｉ ) introduced into the fluid path, which is the value of the independent variable (x) of the regression function, Step (S16) is determined by assuming the following: A step to determine whether a first specified condition is met, wherein the value indicating the predicted occlusion amount (Vpb) is at the current time (t ｃｕｒｒｅｎｔ The amount of the sample fluid (Vin_t) introduced into the fluid path is ) ｃｕｒｒｅｎｔ Step (S18) is determined to be satisfied when it corresponds to the predicted blockage amount (Vpb), and the value of the independent variable (x) of the regression function is the value of the independent variable (x) when the value of the dependent variable (y) is the specified maximum differential pressure. If the first specified condition is met, the sample preparation system (1) is controlled to stop introducing the sample fluid into the fluid path (S20), If the first specified condition is not met, A step of controlling the sample preparation system (1) to continue introducing the sample fluid into the fluid path, The process involves repeating the steps of acquiring system information (S14), determining the regression function (S16), and determining whether or not the first specified condition is met (S18). Methods that include...

2. If the first specified condition is met, the sample preparation system (1) is further controlled so that the rinsing solution flows through the fluid path via the at least one consumable (2a). The method according to claim 1, further comprising:

3. If the first specified condition is met, the sample preparation system (1) is further controlled so that air flows through the fluid path via the at least one consumable (2a). The method according to claim 1, further comprising:

4. If the first specified condition is met, the sample preparation system (1) is further controlled to remove the at least one consumable (2a) from the fluid path. The method according to claim 1, further comprising:

5. The step of further controlling the sample preparation system (1) to remove the at least one consumable (2a) from the fluid path, and then controlling the sample preparation system (1) to provide a new consumable (2a) onto the fluid path. The method according to claim 4, further comprising:

6. The step of controlling the sample preparation system (1) to provide new consumables on the fluid path is performed when a second specified condition is met. The second specified condition may include a condition relating to the amount of remaining sample fluid to be processed by the sample preparation system (1). The method according to claim 5.

7. The aforementioned at least one consumable (2a) is a filter, The aforementioned sample preparation system (1) is The at least one consumable (2a) provided on the fluid path is received, The fluid path is provided with the at least one consumable (2a). The filter handling device (27) is further configured as follows: The step of controlling the sample preparation system (1) to remove the at least one consumable (2a) from the fluid path includes the step of outputting a control signal to the filter handling device (27) to remove the at least one consumable (2a) from the fluid path by receiving the at least one consumable (2a), The step of controlling the sample preparation system (1) to provide the new consumable (2a) on the fluid path includes the step of outputting a control signal to the filter handling device (27) for providing the new consumable (2a) on the fluid path. The method according to claim 5.

8. The sample preparation system (1) further comprises a pump (9) configured to move fluid within the fluid path, The step of controlling the sample preparation system (1) so that the sample fluid flows through the fluid path via the at least one consumable (2a) includes the step of outputting a control signal to the pump (9) to move the sample fluid in the fluid path toward the at least one consumable (2a), The step of controlling the sample preparation system (1) to stop introducing the sample fluid into the fluid path includes the step of outputting a control signal to the pump (9) to stop moving the sample fluid within the fluid path. The method according to claim 1.

9. The sample preparation system (1) comprises at least, The sample fluid is in an open state that allows it to flow towards the at least one consumable (2a) within the fluid path, or A closed state is maintained which prevents the sample fluid from flowing toward the at least one consumable (2a) within the fluid path. The system further comprises a valve (14) configured to switch between the following states: The step of controlling the sample preparation system (1) so that the sample fluid flows through the fluid path through the at least one consumable (2a) includes the step of outputting a control signal to the valve (14) to set the valve (14) to the open state, The step of controlling the sample preparation system (1) to stop introducing the sample fluid into the fluid path includes the step of outputting a control signal to the valve (14) to set the valve (14) to the closed state. The method according to claim 1.

10. The step of determining the regression function is: A step of obtaining one or more values ​​of one or more parameters relating to the sample fluid and / or the sample preparation system (1), A step of generating an equation for determining the amount of blockage as a function of one or more of the one or more parameters, wherein the amount of blockage represents the amount of sample fluid that, when introduced into the at least one consumable, can block the flow of the sample fluid to the at least one consumable (2a). For each of the one or more parameters mentioned above, At present time (t ｃｕｒｒｅｎｔ The amount of the sample fluid (Vin_t) introduced into the fluid path is ) ｃｕｒｒｅｎｔ ) or the total amount of the sample fluid introduced into the fluid path, The amount of blockage obtained using the generated formula, The steps include determining a coefficient that represents the ratio between the two, The steps include determining the regression function using the coefficients determined for each of the one or more parameters, Includes, The one or more of the above parameters are, The cell density of the sample fluid, The turbidity of the sample fluid, The particle size distribution of the sample fluid, The optical density of the sample fluid, The filter resistance index when at least one of the consumables is a filter, When the at least one of the consumables is a filter, the filter surface area, The method according to claim 1, which may include one or more of the following.

11. The step of determining the regression function using the coefficients determined for each of the one or more parameters is: A step of determining a sensitivity coefficient using the coefficients determined for each of the one or more parameters, The steps include selecting one type of regression function to be used as the regression function from among several types of regression functions based on the aforementioned sensitivity coefficient, The method according to claim 10, including the method described in claim 10.

12. The aforementioned sample preparation system (1) is for preparing a sample which is a cell culture medium containing proteins. The sample fluid is the cell culture medium, and the protein contained in the cell culture medium is the target substance. The method according to claim 1.

13. A computer program product comprising computer-readable instructions that, when loaded and executed on a suitable system, causes the system to perform the method according to any one of claims 1 to 12.

14. A control device (50), A processor (502) configured to perform the method described in any one of claims 1 to 12, A storage medium (504) that communicates with the processor, A control device (50) is provided.

15. It is a system, Fluid paths and At least one consumable item (2a), such as a filter, provided on the fluid path, A sample preparation system (1) comprising, The control device (50) described in claim 14, A system that includes these features.

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