Sample preparation system, and method for preparing a sample using the sample preparation system

Abstract

The present invention relates to a sample preparation system and a method for preparing a sample using the sample preparation system.

Description

Technical Field

[0001] The present invention relates to a sample preparation system and a method for preparing a sample using the sample preparation system.

Background Art

[0002] In order to produce a target substance such as a protein by purification, a sample containing the target substance usually undergoes a clarification step before purification to remove contaminants including host cell-derived contaminants such as whole cells, cell fragments, host cell nucleic acids, and host cell proteins, medium-derived contaminants such as proteinaceous and non-proteinaceous medium components, and vector-derived contaminants including vector nucleic acids and viral vectors.

[0003] Clarification and purification are performed in two separate apparatuses. The first apparatus clarifies a sample containing a target substance, and the second separate apparatus uses the clarified output from the first apparatus as an input for purification, which is usually manually supplied to the second apparatus. Therefore, the combination of sample clarification and purification is usually a relatively time-consuming and inefficient process, which may lead to sample degradation.

Summary of the Invention

Problems to be Solved by the Invention

[0004] In view of the above, the technical problem underlying the present invention is to provide a sample preparation system capable of rapidly performing two types of unit operations, such as clarification and purification, in one apparatus in order to improve the efficiency of each process.

Means for Solving the Problems

[0005] The solution to the above technical problem is achieved by embodiments characterized in the claims.

[0006] In particular, in one aspect, the present invention is a sample preparation system, comprising 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 providing direct fluid communication between the first valve and the second valve, a pump configured to move fluid within the sample preparation system, A sample preparation system is provided that includes the above components.

[0007] According to the present invention, the sample preparation system includes a series arrangement of a first means and a second means. In particular, the second means is disposed downstream of the first means within the sample preparation system, and the first means and the second means are integrally connected to each other within the sample preparation system. As used herein, the term "integrally connected" means being disposed within one device (i.e., within the sample preparation system) and not within separate devices. The sample preparation system of the present invention can perform a combination of a first unit operation such as clarification and a second unit operation such as purification in one device, so each method is fast and has significantly improved process efficiency compared to methods performed in two separate devices.

[0008] As used herein, the terms "automated" and "automatically" mean that each step can be performed without human intervention. In contrast, the phrase "manually" as used herein means that the corresponding step is performed by a human.

[0009] The first means of the sample preparation system is not particularly limited as long as it is configured to be connected to the first consumable. According to one embodiment, the first means is configured to be connected to the first consumable and support the first consumable. For example, the first means can be the first filter holder, or the end of the first line, i.e., in this case, the end of the first line that provides fluid communication between the end of the first line and the first valve that supports the first consumable from its downstream side. When the first consumable is supported by the end of the first line from its downstream side, the first consumable is preferably supported on its upstream side by the end of the other hospital upstream of the first means connected to the first means.

[0010] Preferably, the first means is the first filter holder. In one embodiment, the first filter holder includes a first clamp configured to clamp the first consumable on the upper surface (i.e., upstream) and a second clamp configured to clamp the first consumable on the lower surface (i.e., downstream). Thereby, the first consumable can be firmly supported against the internal pressure during operation. According to one embodiment, each of the first clamp and the second clamp includes an O-ring configured to connect the upstream and downstream flow paths of the first means when the O-ring is in a compressed state (i.e., when the clamp is closed). Preferably, one of the first clamp and the second clamp is a fixed clamp, i.e., it does not move, and the other is a movable clamp. Thereby, the first consumable can be clamped by pressing the movable clamp against the fixed clamp. Preferably, the first clamp is a fixed clamp and the second clamp is a movable clamp.

[0011] According to a preferred embodiment, the sample preparation system further includes an input stack configured to hold at least one first consumable. Preferably, the input stack is configured to hold from 1 to 150 first consumables, preferably from 1 to 50 first consumables.

[0012] Furthermore, the second means is not particularly limited as long as it is configured to be connected to the second consumable. According to one embodiment, the second means is configured to be connected to and support the second consumable. For example, the second means is a second filter holder, or corresponds to a portion of a second line that provides fluid communication between the first valve and the second valve.

[0013] When the second means corresponds to a part of the second line, the second line is interrupted between the first valve and the second valve, and the second consumable is disposed at the interruption and supported by a first end of the second line interrupted upstream and a second end of the second line interrupted downstream.

[0014] According to another embodiment where the second means is a second filter holder, the second line is interrupted between the first valve and the second valve, and the second filter holder is disposed at the interruption and connected to a first end of the second line interrupted upstream and a second end of the second line interrupted downstream. In one embodiment, the second filter holder includes a first clamp configured to clamp the second consumable on the upper surface (i.e., upstream) and a second clamp configured to clamp the second consumable on the lower surface (i.e., downstream). Thereby, the second consumable can be firmly supported against the internal pressure during operation. According to one embodiment, each of the first clamp and the second clamp includes an O-ring configured to connect the upstream and downstream flow paths of the second means when the O-ring is in a compressed state (i.e., when the clamp is closed). Preferably, one of the first clamp and the second clamp is a fixed clamp, i.e., it does not move, and the other is a movable clamp. Thereby, the second consumable can be clamped by pressing the movable clamp against the fixed clamp. Preferably, the first clamp is the fixed clamp and the second clamp is the movable clamp.

[0015] In the present invention, a first valve is disposed between a first means and a second means. The first valve is configured to direct fluid to a second line or a third line of the sample preparation system. According to one embodiment, when the fluid is directed to the second line, the flow of fluid to the third line is blocked. According to an alternative embodiment, when the fluid is directed to the third line, the flow of fluid to the second line is blocked. The first valve is not particularly limited. In one embodiment, the first valve can be, for example, a three-way diaphragm valve having three ports, and can be in one of two states that direct flow to one of its two outlets, i.e., one of the second line and the third line. In another embodiment, the first valve is a three-way ball valve.

[0016] Furthermore, a second valve is disposed downstream of the second means. The second valve is configured to direct fluid to a line downstream of the second valve. The second valve is not particularly limited. In one embodiment, the second valve can be, for example, a three-way diaphragm valve having three ports with two inlets connected to the second line and the third line, and can be in one of two states that direct flow from one of the two inlets, i.e., one of the second line and the third line, to the line downstream of the second valve. In another embodiment, the second valve is a three-way ball valve. Preferably, the line is a fourth line described later.

[0017] The pump used in the sample preparation system of the present invention is not particularly limited. The pump is configured to move fluid within the sample preparation system. For example, the pump may be a peristaltic pump, a diaphragm pump, or a piston pump. In one embodiment, the pump is disposed upstream of the first means. Further, the pump may be disposed downstream of an input sample valve described later. Also, the pump may be disposed downstream of a process fluid input valve described later. Thereby, the flow of fluid within the sample preparation system can be easily controlled.

[0018] According to one embodiment of the present invention, the sample preparation system further comprises a consumable handling device configured to provide a first consumable to a first means and connected to the first means configured to receive the first consumable from the first means. The consumable handling device operates in an automated manner. Preferably, the consumable handling device is connected to an input stack, and the consumable handling device automatically receives the first consumable from the first means, provides the first consumable to a waste bin, receives the first consumable from the input stack, and provides the first consumable to the first means, or vice versa. By using the consumable handling device, a new (unused) first consumable can be easily provided to the first means, while at the same time the used first consumable can be easily and automatically removed.

[0019] The consumable handling device is not particularly limited. In one embodiment, the consumable handling device is an index wheel. Preferably, the index wheel has an opening in its in-plane direction at a first position of the index wheel that conforms to the shape of the first consumable. According to one embodiment, the index wheel can be rotated, thereby receiving the first consumable from the input stack and providing it to the first means. Also, the index wheel can be rotated to receive the first consumable from the first means and provide it to the waste bin. In one embodiment, the consumable handling device is a filter handling device.

[0020] In one embodiment of the present invention, the sample preparation system further comprises an input line that provides fluid communication between an input cup disposed upstream of the first means and the first means. According to one embodiment, the input sample valve is disposed downstream of the input cup on the input line and upstream of the first means. Thus, the input sample valve divides the input line into an upstream portion of the input sample valve and a downstream portion of the input sample valve. In this embodiment, the input sample valve is configured to direct fluid to the input line downstream of the input sample valve or to a waste container. According to one embodiment, when fluid is directed to the input line downstream of the input sample valve, the flow of fluid to the waste container is blocked. According to an alternative embodiment, when fluid is directed to the waste container, the flow of fluid to the input line downstream of the input sample valve is blocked. The input sample valve is not particularly limited. In one embodiment, the input sample valve can be, for example, a three-way diaphragm valve having three ports, and can be in one of two states that direct flow to one of two outlets, namely the input line downstream of the input sample valve and the waste container. In another embodiment, the input sample valve is a three-way ball valve. The input cup is configured to hold a fluid, preferably the sample to be prepared. According to one embodiment, the input cup has a volume of from 10 mL to 150 mL, preferably from 20 mL to 80 mL.

[0021] In one embodiment, the sample preparation system further comprises a pipetting robot configured to transfer a fluid, such as the sample to be prepared, from at least one input container to the input cup. In one embodiment, one input container has a volume of from 2 mL to 20 mL. The input container is not particularly limited and can be selected from culture containers, spin tubes, and multi-well plates.

[0022] The pipetting robot that can be used in the present invention is not particularly limited. According to one embodiment, the pipetting robot includes a pipette tip connected to a flow-through pH electrode and a fluid input valve. The fluid input valve is configured to provide, for example, water, sodium hydroxide, and / or a pH neutralization buffer to the pipette tip. Preferably, the pipetting robot further includes a pipette liquid sensor on a pipetting robot line that provides fluid communication between the pipette tip, the pH electrode, and the fluid input valve. The pipette liquid sensor is configured to detect whether or not a liquid is present at its position. Also, the pipetting robot may include a syringe connected to the flow-through pH electrode.

[0023] In one embodiment, the sample preparation system further includes a pipette washing station configured to receive the pipette tip of the pipetting robot. In particular, the pipette washing station may have a wall that surrounds the pipette tip of the pipetting robot while the pipetting robot is placed therein. Further, the pipette washing station may have an outlet at its bottom through which a liquid can come out. Specifically, the liquid can be supplied by at least one of the fluid input valves of the pipetting robot and can be used to wash the pipette tip while the pipetting robot is placed in the pipette washing station. The outlet may be connected to a waste container to which the liquid can be pumped by using a chip washing waste pump.

[0024] Due to the usability of the pipetting robot, the type of the input container is not particularly limited. Therefore, the pipetting robot provides flexibility in the selection of the input container such that its selection depends only on the fluid to be handled, such as the sample to be prepared.

[0025] Preferably, the consumable handling device includes connection means configured to connect the input line and the first line to provide fluid communication between the input line and the first line. Thereby, a through-flow path can be formed between the input line and the first line. When the consumable handling device is an index wheel, the connection means corresponds to an in-plane hole of the index wheel that extends from the first surface of the index wheel to the second surface of the index wheel at the second position of the index wheel. The index wheel can rotate such that a first opening of the hole on the first surface of the index wheel is connected to the input line and a second opening of the hole on the second surface of the index wheel is connected to the first line.

[0026] According to an embodiment of the present invention, the sample preparation system further includes a first liquid sensor disposed upstream of the first means. The first liquid sensor is configured to detect whether or not there is liquid at the position where the first liquid sensor is disposed. Preferably, the first liquid sensor is disposed downstream of the input sample valve and upstream of the first means. According to this configuration, the first liquid sensor can detect, for example, whether or not the input cup is empty.

[0027] In one embodiment, the sample preparation system according to the present invention further includes a second liquid sensor disposed downstream of the first means and upstream of the second means. The second liquid sensor is configured to detect whether or not there is liquid at the position where the second liquid sensor is disposed. Preferably, the second liquid sensor is disposed upstream of the first valve. According to this configuration, based on the detection result of the second liquid sensor, the first valve and the second valve can be switched to guide the fluid through the second line via the second means or through the third line to bypass the second means.

[0028] The sample preparation system according to the present invention may further include a first pressure sensor disposed upstream of the first means. The first pressure sensor is configured to detect the pressure of the sample preparation system at the position where the first pressure sensor is disposed. Preferably, the first pressure sensor is disposed downstream of the pump and upstream of the first means. In one embodiment, the first pressure sensor is disposed downstream of the pump and the first liquid sensor and upstream of the first means. Thereby, the flow of the fluid to the first means can be easily controlled.

[0029] In one embodiment, the sample preparation system of the present invention further includes a second pressure sensor disposed downstream of the first means and upstream of the second means. The second pressure sensor is configured to detect the pressure of the sample preparation system at the position where the second pressure sensor is disposed. Preferably, the second pressure sensor is disposed upstream of the first valve. According to this configuration, based on the measurement result of the second pressure sensor, the first valve and the second valve can be switched to guide the fluid through the second line via the second means or through the third line to bypass the second means.

[0030] In one embodiment, the sample preparation system further includes a fourth line that provides fluid communication between the second valve and an output valve disposed downstream of the second valve. The output valve is configured to direct fluid to an output line that provides fluid communication between the output valve and an output cup, or to a waste container line that provides fluid communication between the output valve and a waste container. According to one embodiment, when the fluid is directed to the output line, the flow of fluid to the waste container line is blocked. According to an alternative embodiment, when the fluid is directed to the waste container line, the flow of fluid to the output line is blocked. The output valve is not particularly limited. In one embodiment, the output valve can be, for example, a three-way diaphragm valve having three ports and can be in one of two states that direct the flow to one of two outlets, namely the output line and the waste container line. In another embodiment, the output valve is a three-way ball valve.

[0031] Preferably, the sample preparation system further comprises an output line providing fluid communication between the output valve and the output cup. The output cup is configured to hold the fluid output by the sample preparation system. In one embodiment, the output cup has a volume of from 10 mL to 150 mL, preferably from 20 mL to 80 mL. According to one embodiment, the sample preparation system comprises a pipetting robot configured to transfer fluid from the output cup to at least one output container. This pipetting robot can be the same as the above-described pipetting robot configured to transfer fluid from at least one input container to the input cup. In one embodiment, one output container has a volume of from 2 mL to 80 mL. The output container is not particularly limited and is selected, for example, from 15 mL centrifuge tubes, 50 mL centrifuge tubes, and multi-well plates.

[0032] Due to the availability of the pipetting robot, the type of output container is not particularly limited. Thus, the pipetting robot provides flexibility in the selection of the output container.

[0033] The sample preparation system according to the present invention may further comprise at least one process fluid input valve disposed upstream of the first means and configured to supply a process fluid to the sample preparation system. The process fluid is not particularly limited and can be selected, for example, from water, phosphate buffered saline (PBS), elution buffer, low pH strip buffer, and sodium hydroxide. The elution buffer is not particularly limited and its selection depends on the substance to be eluted. The elution buffer can be selected, for example, from citrate buffer, glycine / HCl buffer, and acetate buffer. Each of the process fluids may be stored, for example, in a bottle and connected to the process fluid input valve by a line.

[0034] In one embodiment, the sample preparation system according to the present invention may further include a target substance detection sensor disposed downstream of the second means configured to detect the target substance. According to this configuration, the target substance coming out of the first means and / or the second means can be easily detected. In one embodiment, the target substance detection sensor is further disposed downstream of the second valve. Preferably, the target substance detection sensor is further disposed upstream of the output valve. Thereby, according to the detection result of the target substance detection sensor, the fluid can be guided to the output cup or the waste container. Specifically, when the target substance can be detected by the target substance detection sensor, the output valve can be switched to guide the target substance or the fluid containing the target substance to the output cup. When the target substance is not detected by the target substance detection sensor, the output valve can be switched to guide the fluid to the waste container.

[0035] The target substance is not particularly limited. For example, the target substance may be any target biomolecule 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 a preferred embodiment, the target substance is an antibody, more preferably a monoclonal antibody (mAb), or a fragment or derivative thereof, or a nanobody. Examples of monoclonal antibodies are adalimumab, cetuximab, rituximab, infliximab, omalizumab, and denosumab. The target substance can be obtained from, for example, mammalian cells such as "Chinese hamster ovary cells" (CHO cells), HeLa, or human umbilical vein endothelial cells (HUVEC), bacterial cells, or insect cells, media, and cell lines.

[0036] In a preferred embodiment, the target substance detection sensor is a UV sensor. Specifically, by using a UV sensor, a target substance such as a protein can be easily detected.

[0037] The first consumable that can be connected to the first means is not particularly limited. Preferably, the first consumable is a first filter. More preferably, the first filter is a clarification filter. According to one embodiment, the clarification filter is a depth filter and / or a membrane, preferably a depth filter. The clarification filter is configured to separate contaminants from the sample. Preferably, the clarification filter is a cell culture medium clarification filter. The cell culture medium clarification filter is configured to separate cells and contaminants from the sample. In one embodiment, the first consumable is 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 clogging, and is discarded.

[0038] The sample to be prepared contains a plurality of components and is not particularly limited as long as at least one of the plurality of components of the sample is one of the above-described target substances. The additional (impurity) components (contaminants) are not particularly limited and may depend on the preparation conditions of the target substance. Examples of additional components are aggregates, host cell proteins, deoxyribonucleic acid, and fragments and charge variants thereof.

[0039] The source of the sample is not particularly limited. For example, the sample can be obtained by applying any biological, biochemical, chemical, or pharmaceutical method. Thereby, the sample 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.

[0040] The second consumable that can be connected to the second means is not particularly limited. Preferably, the second consumable is a second filter or a chromatography column. More preferably, the second consumable is a purification filter or a purification chromatography column. According to one embodiment, the purification filter is a membrane. In one embodiment, the purification chromatography column contains functionalized beads. Preferably, the second consumable is configured to bind and elute the target substance. In one embodiment, the second consumable is a protein purification filter or a protein purification chromatography column, more preferably a protein purification filter. Preferably, the protein purification filter or the protein purification chromatography column is configured to bind and elute a protein, preferably an antibody. In one embodiment, the second consumable is a reusable consumable. As used herein, the term "reusable consumable" means that the consumable can be used for several sample preparations before it has to be discarded. For example, the reusable consumable needs to be replaced after performing 50 to 150 operations. According to one embodiment, the purification filter or the purification chromatography column each contains a binding compound that enables binding of the target substance within the purification filter or the purification chromatography column. For example, the binding compound is immobilized on / in the filter matrix of the purification filter. The binding compound may be, for example, Protein A that can bind to the target substance at a pH of 7 or higher. Then, an elution buffer having a pH of less than 7 can be used to elute the target substance.

[0041] In one embodiment, the sample preparation system further comprises a user interface touch screen. The user interface touch screen enables easy control and operation of the sample preparation system.

[0042] The sample preparation system of the present invention may further include a pH sensor disposed downstream of the second means. The pH sensor is configured to measure the pH of the liquid at the position of the pH sensor. Preferably, the pH sensor is further disposed downstream of the second valve and upstream of the output valve. According to one embodiment, the sample preparation system further includes a pH adjustment input line that provides fluid communication between the pH adjuster reservoir and a second line upstream of the second means. Thereby, the pH in the second consumable that can be connected to the second means can be easily controlled. In addition, the sample preparation system may further include an elution fraction line that provides fluid communication between the elution fraction reservoir and a second line downstream of the second means.

[0043] The material of the line of the present invention is not particularly limited. Preferably, the material can be selected from plastics such as polyvinyl chloride, polypropylene, polyethylene, polytetrafluoroethylene, and fluorinated ethylene propylene. Thereby, the line can provide flexibility and chemical resistance.

[0044] The sample preparation system according to the present invention may include two or more series arrangements of the first means and the second means. In particular, each of these series arrangements may have the same configuration as the series arrangement described above. When two or more series arrangements are present in the sample preparation system, the series arrangements are arranged parallel to each other, and each of the series arrangements has its own input cup and output cup to enable independent operation.

[0045] The sample preparation system of the present invention enables two unit operations, such as clarification and purification, to be performed within one device so that each method has high speed and improved process efficiency compared to methods using separate devices. In particular, target substances contained in more than 20 input containers containing unfiltered samples can be clarified and purified, for example, in less than 2 hours. Thus, the sample preparation system according to the present invention can provide a fast method for clarifying and purifying samples so as to avoid sample degradation. Further, the automated operation of the sample preparation system can reduce the need to perform each process manually. Also, the sample preparation system can generate a clear (i.e., representative) output sample containing a target substance that can be directly subjected to further analysis via various assays.

[0046] According to another aspect, the present invention is a method for preparing a sample using a sample preparation system, the method comprising: (i) clarifying the sample by passing the sample through a first consumable connected to a first means for supplying a clarified sample, the first consumable being a clarification filter (hereinafter referred to as "clarification process (i)"), and (ii) purifying the sample by passing the sample through a second consumable connected to a second means for supplying a purified sample, the second consumable being a purification filter or a purification chromatography column (hereinafter referred to as "purification process (ii)") providing a method comprising at least one of.

[0047] The present invention provides a method for preparing a sample using a sample preparation system, the method comprising a clarification process (i), a purification process (ii), or both a clarification process (i) and a purification process (ii).

[0048] The following detailed description of the clarification process (i) relates to a method for preparing a sample using a sample preparation system, the method including the clarification process (i) but not including the purification process (ii). The following detailed description of the purification process (ii) relates to a method for preparing a sample using a sample preparation system, the method including the purification process (ii) but not including the clarification process (i). Details of a method for preparing a sample using a sample preparation system, the method including both the clarification process (i) and the purification process (ii), will be presented later.

[0049] Any definition of a feature made in the context of the sample preparation system applies to each feature of the method for preparing a sample using the sample preparation system, unless otherwise specified.

[0050] According to a preferred embodiment of the method for preparing a sample of the present invention, 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 the purification chromatography column is a protein purification chromatography column. Each method using the sample preparation system according to the present invention can clarify and purify the sample more efficiently and quickly compared to a method in which clarification and purification are performed in two separate apparatuses. Further, the method of the present invention can generate a clear (i.e., representative) output sample containing a target substance such as a protein, preferably an antibody, which can be directly subjected to further analysis via various assays.

[0051] Clarification process (i) According to one embodiment, at the beginning of the clarification process (i), a clarification filter is provided in the first means, and the clarification filter is new, i.e., 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.

[0052] In another embodiment, the clarification filter may already be connected to the first means at the start of the clarification process (i). In both embodiments, it may be necessary to replace the clarification filter during the clarification process (i) to avoid clogging of the clarification filter and product loss. According to one embodiment, the clarification filter is automatically discharged from the first means and automatically exchanged with a new clarification filter. Preferably, based on the measured backpressure of the clarification filter measured by the first pressure sensor, or the pressure difference between the first pressure sensor and the second pressure sensor, and the pumping speed, a decision can be made to automatically discharge the clarification filter from the first means and automatically exchange it with a new clarification filter.

[0053] The clarification filter is then 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.

[0054] Furthermore, an air pressure test can be performed on the sample preparation system and the clarification filter before the sample to be clarified is pumped through 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 pumps air into the sample preparation system to reach a pre-set pressure. According to one embodiment, 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 such that the pressure is released to the waste container via the third line.

[0055] After an optional air pressure test, a sample to be clarified can be supplied to the input cup of the sample preparation system (i.e., "step (c4)a"). Alternatively, a sample to be clarified can also be supplied to the input cup of the sample preparation system before any of steps (c1) to (c3) (i.e., "step (c4)b"). Preferably, the pipetting robot can supply the sample to be clarified to the input cup by transferring the sample to be clarified from at least one input container to the input cup of the sample preparation system.

[0056] Next, the sample to be clarified can be pumped from the input cup to the clarification filter (i.e., "step (c5)"). According to one embodiment of the above step (c5), the sample to be clarified passes through at least an input sample valve, a first liquid sensor, and a first pressure sensor on the way from the input cup to the clarification filter.

[0057] Furthermore, the sample is clarified by passing it through a clarification filter connected to the first means (hereinafter referred to as "step (c6)").

[0058] Next, the clarified sample can be induced into the output cup (hereinafter referred to as "step (c7)"). According to one embodiment of the above step (c7), the clarified sample passes through at least a second pressure sensor, a second liquid sensor, a third line, a target substance detection sensor, and an output valve from the downstream side of the clarification filter to the output cup.

[0059] When the input cup is empty, the first liquid sensor can detect that the sample to be clarified is gone (i.e., "step (c8)"). Specifically, the first liquid sensor can detect whether the sample to be clarified previously supplied to the input cup has already passed the position where the first liquid sensor is disposed.

[0060] 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 by a process fluid input valve disposed upstream of the first means. Thereby, the clarified sample still present upstream of the first means can be induced to the first means, 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.

[0061] 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 by a process fluid input valve disposed upstream of the first means. 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. The clarified sample thus generated can be induced to the output cup. By applying step (c10), the product recovery rate can be increased. The air pressure used is, for example, in the range of 0.5 bar to 2 bar. Thereby, the air pressure is 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. After a waiting time of, for example, 0 seconds to 5 seconds, the air pressure is released by reversing the pump. The measurement of the bubble point is well known to those skilled in the art. For example, the bubble point can be measured according to ISO 2942 or ASTM F316-03.

[0062] Next, the clarification filter can be discharged from the first means (i.e., "step (c11)"). When the first means is the first filter holder and the clarification filter is clamped by the first clamp and the second clamp, the clarification filter is unclamped before it is discharged. The discharged clarification filter can be received by the consumable handling device and supplied to the waste bin.

[0063] Furthermore, according to one embodiment, the connection means included in the consumable handling device connects the input line and the first line (i.e., "step (c12)") to provide fluid communication between the input line and the first line so that the liquid path is closed. This can be achieved, for example, by using an index wheel as the consumable handling device and rotating the index wheel to the second position of the index wheel where a through-fluid passage that can then be connected to the input line and the first line is arranged.

[0064] 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 by pumping air (i.e., "step (c13)").

[0065] In one embodiment of the present invention, 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.

[0066] Optionally, the sample preparation system can be washed by pumping a cleaning liquid such as a buffer solution, sodium hydroxide, and / or water from the process fluid input valve arranged upstream of the first means through the sample preparation system to the output cup and then reversely from the output cup to the input cup (i.e., "step (c15)"). Then, the resulting liquid can be induced into the waste container using a waste pump.

[0067] Purification process (ii) The purification filter or purification chromatography column used in the purification process (ii) preferably already exists within the sample preparation system before the purification process (ii) is initiated. The purification filter or purification chromatography column can be provided manually, for example, in the sample preparation system. In another embodiment, the purification filter or purification chromatography column can be provided in the sample preparation system in an automated manner.

[0068] According to a preferred embodiment, the purification filter or purification chromatography column is in an equilibrium state. As used herein, the term "equilibrium state" means that the purification filter or purification chromatography column is in a state where it can bind the target substance. This state can be achieved, for example, by setting the purification filter or purification chromatography column to a specific pH such as 7 or higher while containing the binding compound. If the purification filter or purification chromatography column is not in an equilibrium state, an equilibrium buffer such as PBS can be pumped through the purification filter or purification chromatography column. This step can be performed at any point in the purification process (ii) before step (p3) described below.

[0069] Furthermore, according to one embodiment, in order to avoid air lock blockage of the purification filter or the purification chromatography column, excessive air can be removed from the upstream side of the purification filter or the purification chromatography column. In particular, a buffer solution such as PBS can be pumped into the output cup, and then the pump is reversed to draw the buffer solution through the purification filter or the purification chromatography column from its outlet to its inlet while monitoring and controlling the negative pressure at the inlet of the purification filter or the purification chromatography column by the second pressure sensor. In one embodiment, the pressure is controlled to be an absolute pressure of 0.4 bar or more. Then, after the equilibration step and / or the step of removing excessive air from the purification filter or the purification chromatography column, the lines of the sample preparation system except for the input cup, the output cup, and the second line can be emptied.

[0070] According to one embodiment, at the beginning of the purification process (i), a through-flow path is provided in the first means that connects the line upstream of the first means and the first line of the sample preparation system (i.e., "step (p1)"). Preferably, the line upstream of the first means is the input line. If a clarification filter is present in the first means, the clarification filter is discharged before the through-flow path is provided. In one embodiment of the present invention, the connection means included in the consumable handling device connects the input line and the first line to provide fluid communication between the input line and the first line, thereby providing a through-flow path in the first means. This can be achieved, for example, by using an index wheel as the consumable handling device and rotating the index wheel to the second position of the index wheel where a through-fluid passage that can then be connected to the input line and the first line is arranged.

[0071] Optionally, if the sample preparation system comprises a target substance detector, the detector may be zeroed before the sample to be purified is supplied to the purification filter or purification chromatography column (i.e., "step (p2)"). For example, a buffer such as PBS can be supplied to the sample preparation system by a process fluid input valve disposed upstream of the first means. Preferably, the buffer is pumped through the third line of the sample preparation system rather than the second line of the sample preparation system.

[0072] Next, the sample to be purified can be supplied to the input cup of the sample preparation system (i.e., "step (p3)"). Preferably, the pipetting robot can supply the sample to be purified to the input cup by transferring the sample to be purified from at least one input container to the input cup of the sample preparation system.

[0073] Thereafter, the sample to be purified can be pumped in the direction of the purification filter or the purification chromatography column (i.e., "step (p4)"). According to one embodiment, while the sample to be purified is being pumped in the direction of the purification filter or the purification chromatography column, the first valve and the second valve are switched to bypass the second means to avoid pumping excessive air into the purification filter or the purification chromatography column that may lead to performance degradation and ultimately air lock blockage of the purification filter or the purification chromatography column. In one embodiment, when the sample to be purified is detected by the first liquid sensor, the first valve and the second valve are switched to direct the sample to be purified through the purification filter or the purification chromatography column. Preferably, when the sample to be purified is detected by the second liquid sensor, the first valve and the second valve are switched to direct the sample to be purified through the purification filter or the purification chromatography column. Thereby, even less air is pumped into the purification filter or the purification chromatography column. Optionally, after detection of the sample to be purified through the first liquid sensor or the second liquid sensor, a predetermined volume of the sample to be purified may be pumped before switching the first valve and the second valve, and thus even less air is pumped into the purification filter or the purification chromatography column, or no air is pumped in.

[0074] When the input cup is empty, the first liquid sensor can detect that the sample to be purified is gone (i.e., "step (p5)"). Specifically, the first liquid sensor can detect whether the sample to be purified previously supplied to the input cup has already passed the position where the first liquid sensor is disposed.

[0075] Subsequently, the rinse step can be automatically executed (i.e., "step (p6)"). In particular, a process fluid input valve disposed upstream of the first means can supply a rinse liquid such as PBS to the sample preparation system. Thereby, the sample to be purified that still exists upstream of the second means can be induced to the second means and purified by passing it through a purification filter or a purification chromatography column.

[0076] Subsequently, according to one embodiment, the target substance detection sensor included in the sample preparation system can be set to zero after the rinse step (i.e., "step (p7)"). For example, a buffer solution such as an elution buffer can be supplied to the sample preparation system by a process fluid input valve disposed upstream of the first means. Preferably, the buffer solution is pumped through the third line of the sample preparation system rather than the second line of the sample preparation system.

[0077] Optionally, after step (p6) and / or step (p7), any liquid still contained at least in the third line of the sample preparation system can be discharged (i.e., "step (p8)"). In one embodiment of step (p8), any liquid still present in the input line, the first line, and / or the fourth line of the sample preparation system can be further discharged. Thereby, air can be introduced into the third line that can be used for pushing into the sample output cup in a later step.

[0078] Next, according to one embodiment, an elution step is performed (i.e., "step (p9)"). Preferably, while a buffer solution such as an elution buffer is being pumped in the direction of the purification filter or the purification chromatography column, the first valve and the second valve are switched to bypass the second means in order to avoid pumping excessive air into the purification filter or the purification chromatography column that would result in performance degradation and ultimately an airlock blockage of the purification filter or the purification chromatography column. In one embodiment, when a buffer solution such as an elution buffer is detected by the first liquid sensor, the first valve and the second valve are switched to direct the buffer solution through the second line via the purification filter or the purification chromatography column. Preferably, when a buffer solution such as an elution buffer is detected by the second liquid sensor, the first valve and the second valve are switched to direct the buffer solution through the second line via the purification filter or the purification chromatography column. Thereby, even less air is pumped into the purification filter or the purification chromatography column. Optionally, after detection of a buffer solution such as an elution buffer through the first liquid sensor or the second liquid sensor, a predetermined volume of the buffer solution may be pumped before switching the first valve and the second valve, and thus even less air is pumped into the purification filter or the purification chromatography column, or no air is pumped. A buffer solution can be supplied to the sample preparation system by a process fluid input valve disposed upstream of the first means. In one embodiment of step (p9), the elution buffer may be a mixture of a first elution buffer and a second elution buffer, the first elution buffer can be supplied to the sample preparation system by a first process fluid input valve disposed upstream of the first means, and the second elution buffer can be supplied to the sample preparation system by a second process fluid input valve disposed upstream of the first means. Thereby, gradient elution can be performed by the controlled use of the first and second process fluid input valves.

[0079] Thereafter, the purified sample can be collected in the output cup (i.e., "step (p10)"). In one embodiment, the purified sample corresponds to a fraction containing the target substance eluted from the purification filter or the purification chromatography column. Specifically, the fraction can be eluted using an elution buffer in step (p9). According to one embodiment, the concentration of the target substance in the fraction is higher than 0.4 g / L, preferably higher than 0.5 g / L. Thus, the fraction is suitable for direct analysis via various subsequent assays.

[0080] The purified sample can be collected in different ways in step (p10) above. According to one embodiment, a target substance detection sensor can be used to detect when the target substance passes through the sensor. In another embodiment, the sample may be purified using a predetermined elution volume. In both embodiments, if a rinse step (step (p6)) is performed before step (p10), most of the first liquid coming out of the purification filter or the purification chromatography column is the buffer used in the rinse step and contains little target substance, and thus is preferably diverted to the waste container. When the target substance detection sensor detects that the target substance or an appropriate amount of the predetermined elution volume has eluted, the output valve is switched to divert the purified sample to the output cup. In a preferred embodiment, the target substance detection sensor is a UV sensor. Thereafter, the total eluted target substance can be calculated using the output from the UV sensor, the volume of the elution buffer pumped from the system, the optical path length of the target substance such as protein, and the extinction coefficient.

[0081] In one embodiment, after sufficient buffer such as the elution buffer is pumped in the direction of the purification filter or the purification chromatography column, the first valve and the second valve can be optionally switched so that the air that may be present in the third line of the sample preparation system provided in the previous step (i.e., step (p8)) can be used to divert the purified sample still present in the output line to the output cup (i.e., "step (p11)").

[0082] Preferably, the output sample present in the output cup can be transferred to at least one output container (i.e., "step (p12)"). This step can be performed, for example, by a pipetting robot.

[0083] Optionally, the sample preparation system can be washed by pumping a cleaning liquid such as a buffer, sodium hydroxide, and / or water from the process fluid input valve disposed upstream of the first means through the sample preparation system to the output cup and then reversely from the output cup to the input cup (i.e., "step (p13)"). Then, the resulting liquid can be directed to a waste container using a waste pump.

[0084] According to one embodiment, the purification filter or purification chromatography column is then removed from the sample preparation system if it is at the end of its useful life (i.e., "step (p14)").

[0085] Combination of clarification process (i) and purification process (ii) In a method (hereinafter referred to as the "composite method") for conditioning a sample using a sample preparation system including a clarification process (i) and a purification process (ii), the clarification process (i) (i.e., step (i)) is performed before the purification process (ii) (i.e., step (ii)).

[0086] The purification filter or purification chromatography column used in the purification process (ii) of the composite method is preferably already present in the sample preparation system before the clarification process (i) is initiated. The purification filter or purification chromatography column can be provided manually in the sample preparation system, for example. In another embodiment, the purification filter or purification chromatography column can be provided in the sample preparation system in an automated manner.

[0087] According to a preferred embodiment, the purification filter or the purification chromatography column is in an equilibrium state. As used herein, the term "equilibrium state" means that the purification filter or the purification chromatography column is in a state where it can bind the target substance. This state can be achieved, for example, by adjusting the purification filter or the purification chromatography column to a specific pH, such as 7 or higher, while containing the binding compound. If the purification filter or the purification chromatography column is not in an equilibrium state, an equilibration buffer such as PBS can be pumped through the purification filter or the purification chromatography column. This step can be performed at any point in the conjugation method prior to step (4)a or step (4)b described below.

[0088] Furthermore, according to one embodiment, in order to avoid air lock blockage of the purification filter or the purification chromatography column, excess air can be removed from the upstream side of the purification filter or the purification chromatography column. In particular, a buffer such as PBS can be pumped into the output cup, and then the pump can be reversed to draw the buffer through the purification filter or the purification chromatography column from its outlet to its inlet while monitoring and controlling the negative pressure at the inlet of the purification filter or the purification chromatography column by a second pressure sensor. In one embodiment, the pressure is controlled to be an absolute pressure of 0.4 bar or higher. Then, after the equilibration step and / or the step of removing excess air from the purification filter or the purification chromatography column, the lines of the sample preparation system except for the input cup, the output cup, and the second line can be emptied.

[0089] According to one embodiment, at the beginning of the clarification process (i) of the conjugation method, a clarification filter is provided in the first means, and the clarification filter is new, i.e., unused (hereinafter referred to as "step (1)"). In said step (1), preferably, the clarification filter is provided by a consumable handling device from an input stack.

[0090] In another embodiment, the clarification filter may already be connected to the first means at the start of the clarification process (i). In both embodiments, it may be necessary to replace the clarification filter during the clarification process (i) to avoid clogging of the clarification filter and product loss. According to one embodiment, the clarification filter is automatically discharged from the first means and automatically exchanged with a new clarification filter. Preferably, based on the measured backpressure of the clarification filter measured by the first pressure sensor, or the pressure difference between the first pressure sensor and the second pressure sensor, and the pumping speed, a decision can be made to automatically discharge the clarification filter from the first means and automatically exchange it with a new clarification filter.

[0091] The clarification filter is then preferably clamped (i.e., "step (2)") such that the upper and lower surfaces of the clarification filter are supported against the internal pressure during filtration.

[0092] Furthermore, an air pressure test can be performed on the sample preparation system and the clarification filter before the sample to be clarified and purified is pumped through the clarification filter (i.e., "step (3)"). In particular, in said step (3), the first valve and the second valve are set to close the flow path, and the pump pumps air into the sample preparation system so as to reach a preset pressure. According to one embodiment, the preset 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 such that the pressure is released to the waste container via the third line.

[0093] After an optional air pressure test, a clarified and purified sample can be supplied to the input cup of the sample preparation system (i.e., "step (4)a"). Alternatively, it is also possible to supply a clarified and purified sample to the input cup of the sample preparation system before any of steps (1) to (3) (i.e., "step (4)b"). Preferably, the pipetting robot can supply the clarified and purified sample to the input cup by transferring the clarified and purified sample from at least one input container to the input cup of the sample preparation system.

[0094] Next, the clarified and purified sample can be pumped from the input cup to the clarification filter (i.e., "step (5)"). According to one embodiment of the above step (5), the clarified and purified sample passes through at least the input sample valve, the first liquid sensor, and the first pressure sensor on the way from the input cup to the clarification filter.

[0095] Furthermore, the sample is clarified by passing it through a clarification filter connected to the first means (hereinafter referred to as "step (6)").

[0096] Next, the clarified sample immediately flows from the clarification filter to the purification filter or the purification chromatography column (i.e., "step (7)"). Therefore, the clarified sample coming out of the clarification filter is not directly guided to the output cup as described for one embodiment of the method for adjusting the sample using the sample preparation system. This method includes the clarification process (i) but does not include the purification process (ii), and instead is immediately guided to the purification filter or the purification chromatography column.

[0097] When the input cup is empty, the first liquid sensor can detect that there is no sample to be clarified and purified (i.e., "Step (8)"). Specifically, the first liquid sensor can detect whether the sample to be clarified and purified previously supplied to the input cup has already passed the position where the first liquid sensor is disposed.

[0098] After Step (8), a rinse step can be automatically executed (i.e., "Step (9)"). In particular, a rinse liquid such as PBS can be supplied to the sample preparation system by a process fluid input valve disposed upstream of the first means. Thereby, the sample to be clarified and purified still present upstream of the first means can be induced to the first means, 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 purification filter or the purification chromatography column. Thereby, the clarified sample that still needs to be purified and is present upstream of the second means can be induced to the second means and purified by passing it through the purification filter or the purification chromatography column.

[0099] After step (8) and / or step (9), preferably, a step of pumping air is performed (i.e., "step (10)"). Specifically, air can be supplied to the sample preparation system by a process fluid input valve disposed upstream of the first means. The air pushes the clarified and purified sample or rinse liquid still present in the upper flow field of the clarification filter, and the sample or rinse liquid that is already partially clarified but not yet purified and still present in the clarification filter, and passes through the clarification filter. The clarified sample thus produced can be directed to a purification filter or a purification chromatography column. Further, by the step of pumping air, the clarified sample that needs to be purified can be pushed and passed through a purification filter or a purification chromatography column. By applying step (10), the product recovery rate can be increased. The air pressure used is, for example, in the range of 0.5 bar to 2 bar. Thereby, the air pressure is below the bubble point of the clarification filter so as to avoid the formation of air bubbles. After a waiting time of, for example, 0 seconds to 5 seconds, the air pressure is released by reversing the pump.

[0100] According to one embodiment of the composite method, the method is, between step (i) and step (ii), that is, at least after the above step (6), in the following order, (a) a step of discharging the clarification filter; (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; (c) a 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; and further includes.

[0101] In particular, in the above step (a), the clarification filter is discharged from the first means (i.e., "step (11)"). When the first means is the first filter holder and the clarification filter is clamped by the first clamp and the second clamp, the clarification filter is unclamped before it is discharged. The discharged clarification filter can be received by the consumable handling device and supplied to the waste bin.

[0102] Furthermore, according to one embodiment, the through-flow path in step (b) above can be provided using connection means included in the consumable handling device that connects the input line and the first line to provide fluid communication between the input line and the first line so that the liquid path is closed (i.e., "step (12)"). This can be achieved, for example, by using an index wheel as the consumable handling device and rotating the index wheel to the second position of the index wheel where a through-fluid passage that can then be connected to the input line and the first line is arranged.

[0103] Then, in order to remove the sample from the line upstream of the first means according to step (c) above, a rinse step can be performed (i.e., "step (13)"). In particular, a rinse liquid such as PBS can be supplied to the sample preparation system by a process fluid input valve arranged upstream of the first means. Preferably, in this embodiment of step (c), the first valve and the second valve can be switched so as to bypass the second means so that the rinse liquid with the unclarified sample can be induced through the through-flow path provided in step (b), the first line and the third line of the sample preparation system. Thereby, all unclarified samples still present upstream of the first means of the sample preparation system can be removed to avoid contamination and blockage of the purification filter or the purification chromatography column. Furthermore, in one embodiment, step (13) may also optionally be used to zero the target substance detection sensor.

[0104] Thereafter, the first valve and the second valve can be switched (i.e., "step (14)") to direct the rinse liquid through a second line via a purification filter or a purification chromatography column. Thereby, the clarified sample that still needs to be purified and that is present downstream of the first valve and upstream of the second means can be directed to the second means and purified by passing it through a purification filter or a purification chromatography column.

[0105] In one embodiment, after step (13) and / or step (14), any liquid still contained in at least a third line of the sample preparation system can be drained (i.e., "step (15)"). In one embodiment of step (15), any liquid still present in the input line and / or the first line of the sample preparation system can be further drained. Thereby, air can be introduced into the third line that can be used to push into the sample output cup in a later step.

[0106] Next, according to one embodiment, an elution step is performed (i.e., "step (16)"). Preferably, while a buffer solution such as an elution buffer is being pumped in the direction of the purification filter or the purification chromatography column, the first valve and the second valve are switched to bypass the second means in order to avoid pumping excessive air into the purification filter or the purification chromatography column that would result in performance degradation and ultimately an airlock blockage of the purification filter or the purification chromatography column. In one embodiment, when a buffer solution such as an elution buffer is detected by the first liquid sensor, the first valve and the second valve are switched to direct the buffer solution through the second line via the purification filter or the purification chromatography column. Preferably, when a buffer solution such as an elution buffer is detected by the second liquid sensor, the first valve and the second valve are switched to direct the buffer solution through the second line via the purification filter or the purification chromatography column. Thereby, even less air is pumped into the purification filter or the purification chromatography column. Optionally, after detection of a buffer solution such as an elution buffer through the first liquid sensor or the second liquid sensor, a predetermined volume of the buffer solution may be pumped before switching the first valve and the second valve, and thus even less air is pumped into the purification filter or the purification chromatography column, or no air is pumped. A buffer solution can be supplied to the sample preparation system by a process fluid input valve disposed upstream of the first means. In one embodiment of step (16), the elution buffer may be a mixture of a first elution buffer and a second elution buffer, the first elution buffer can be supplied to the sample preparation system by a first process fluid input valve disposed upstream of the first means, and the second elution buffer can be supplied to the sample preparation system by a second process fluid input valve disposed upstream of the first means. Thereby, gradient elution can be performed by the controlled use of the first and second process fluid input valves.

[0107] Thereafter, the clarified and purified sample can be collected in an output cup (i.e., "step (17)"). In one embodiment, the clarified and purified sample corresponds to a fraction containing the target substance eluted from a purification filter or a purification chromatography column. Specifically, the fraction can be eluted using an elution buffer in step (16). According to one embodiment, the concentration of the target substance in the fraction is higher than 0.4 g / L, preferably higher than 0.5 g / L. Therefore, the fraction is suitable for direct analysis via various subsequent assays.

[0108] The clarified and purified sample can be collected in different ways in step (17) above. According to one embodiment, a target substance detection sensor can be used to detect when the target substance passes through the sensor. In another embodiment, the collection of the clarified and purified sample may be based on a step using a predetermined elution volume. In both embodiments, if a rinse step (step (13)) is performed before step (17), most of the first liquid coming out of the purification filter or the purification chromatography column is the buffer used in the rinse step, contains little target substance, and is therefore preferably diverted to a waste container. When the target substance detection sensor detects that the target substance or an appropriate amount of the predetermined elution volume has eluted, the output valve is switched to divert the clarified and purified sample to the output cup. In a preferred embodiment, the target substance detection sensor is a UV sensor. Thereafter, the total eluted target substance can be calculated using the output from the UV sensor, the volume of the elution buffer pumped from the system, the optical path length of the target substance such as protein, and the extinction coefficient.

[0109] In one embodiment, after sufficient buffer such as elution buffer is pumped in the direction of the purification filter or the purification chromatography column, the first valve and the second valve can be optionally switched (i.e., "step (18)") so that air that may be present in the third line of the sample preparation system provided in the previous step (i.e., step (15)) can be used to direct the clarified and purified sample still present in the output line to the output cup.

[0110] Preferably, the output sample present in the output cup can be transferred to at least one output container (i.e., "step (19)"). This step can be performed, for example, by a pipetting robot.

[0111] Optionally, the sample preparation system can be washed by pumping a cleaning liquid such as buffer, sodium hydroxide, and / or water from the process fluid input valve disposed upstream of the first means through the sample preparation system to the output cup and then back from the output cup to the input cup (i.e., "step (20)"). Then, the resulting liquid can be directed to a waste container using a waste pump.

[0112] According to one embodiment, the purification filter or the purification chromatography column is then removed from the sample preparation system when it is at the end of its useful life (i.e., "step (21)").

Brief Description of the Drawings

[0113]

Figure 1

Figure 2

Figure 3

Figure 4

Description of Symbols

[0114] 1 Sample preparation system 2 First means 2a First consumable 3 Second means 3a Second consumable 4 First valve 5 Second valve 6 First line 7 Second line 8 Third line 9 Pump 10a First clamp 10b Second clamp 11 Input stack 12 Input line 13 Input cup 14 Input sample valve 15 First liquid sensor 16 Second liquid sensor 17 First pressure sensor 18 Second pressure sensor 19 Fourth line 20 Output valve 21 Target substance detection sensor 22 Output line 23 Output cup 24 Process fluid input valve 25 Waste pump 26 Waste bin 27 Consumable handling device 28 Opening 29 Hole 30 Pipetting robot 31 Pipette tip 32 pH electrode 33 Fluid input valve 34 Pipette liquid sensor 35 Pipetting robot line 36 Syringe 37 Pipette washing station 38 Wall 39 Outlet 40 Chip washing waste pump 41 Storage container 42 Input container 43 Output container

Claims

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 positioned between the first means and the second means, A second valve positioned downstream of the second means, A first line provides fluid communication between the first means and the first valve, A second line provides fluid communication between the first valve and the second valve via the second means, A third line, configured to bypass the second means, provides direct fluid communication between the first valve and the second valve, A pump configured to move fluid within the sample preparation system, A sample preparation system equipped with the following features.

2. The first means is connected to the first consumable and configured to support the first consumable, The sample preparation system according to claim 1.

3. The second means is connected to the second consumable and configured to support the second consumable, The sample preparation system according to claim 1.

4. The first consumable is the first filter. The sample preparation system according to claim 1.

5. The second consumable is a second filter or chromatography column. The sample preparation system according to claim 1.

6. The first means is a first filter holder. The sample preparation system according to claim 1.

7. A consumable handling device connected to the first means, which is configured to supply the first consumable to the first means and to receive the first consumable from the first means. The sample preparation system according to claim 1, further comprising:

8. The system further comprises an input line that provides fluid communication between an input cup located upstream of the first means and the first means, The consumable handling device includes connecting means configured to connect the input line and the first line in order to provide fluid communication between the input line and the first line. The sample preparation system according to claim 7.

9. The input sample valve is further located downstream of the input cup on the input line and upstream of the first means, The sample preparation system according to claim 8.

10. The system further comprises a target substance detection sensor located downstream of the second means configured to detect a target substance, The sample preparation system according to claim 1.

11. A method for preparing a sample using the sample preparation system described in any one of claims 1 to 10, The aforementioned method, (i) a step of clarifying a sample by passing it through a first consumable connected to the first means in order to supply a clarified sample, wherein the first consumable is a clarification filter, and (ii) A step of purifying the sample by passing it through a second consumable connected to the second means in order to supply the purified sample, wherein the second consumable is a purification filter or a purification chromatography column. A method that includes at least one of the following.

12. The method comprises steps (i) and (ii), where step (i) is performed before step (ii), and the clarified sample flows immediately from the clarification filter to the purification filter or the purification chromatography column. A method for preparing the sample according to claim 11.

13. The above method involves the following steps between step (i) and step (ii): (a) A step of discharging the clarification filter, (b) Providing a through-channel within the first means that connects the upstream line of the first means and the first line of the sample preparation system, (c) A step of removing the sample from the line upstream of the first means by guiding the sample through the through-flow channel, the first line, and the third line of the sample preparation system, A method for preparing the sample according to claim 12, further comprising:

14. The aforementioned sample is a cell culture medium containing protein, The clarification filter is a cell culture medium clarification filter. The purification filter is a protein purification filter or a protein purification chromatography column. A method for preparing the sample according to claim 11.

15. In step (i), the clarification filter is automatically discharged from the first means and automatically replaced with a new clarification filter. A method for preparing the sample according to claim 11.

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