Method for controlling liquid transport in flow path of biomolecule analyzer using computer, and biomolecule purification system

Abstract

To prevent air from exceeding a purification membrane even in the absence of an additional flow path structure, the present disclosure proposes a method for controlling liquid transfer in a flow path of a biomolecule analyser using a computer, the biomolecular analyser having a first chamber containing a first liquid, a second chamber containing a second liquid, a membrane chamber having a purification membrane, and a waste liquid chamber, the method including: performing control, by computer, so as to transport the second liquid until at least beyond a confluence of a first flow path leading from the first chamber to the waste liquid chamber and a second flow path extending from the second chamber, and draining a fluid different from the first and second liquids from the second flow path; performing control, by computer, so as to transport the first liquid from the first chamber to the waste liquid chamber via the membrane chamber; and performing control, by computer, so as to transport the second liquid in the second chamber from the second chamber to the membrane chamber (see fig. 6).

Description

Method for controlling liquid transport in a flow channel of a biomolecule analyzer by a computer, and biomolecule purification system

[0001] The present disclosure relates to a method for controlling liquid transport in a flow channel of a biomolecule analyzer by a computer, and a biomolecule purification system.

[0002] When analyzing genes, for example, preprocessing involves dissolving the sample, purifying and amplifying the nucleic acid, and then detecting the amplified product. This process involves the risk of contamination and the need for complex sample preparation. For this reason, the conventional procedure has been to send samples to an environment with well-equipped experimental facilities, such as a research lab, where inspectors with specialized knowledge and skills prepare the samples, measure them, and then analyze the data. However, problems include the time required to transport the samples and the large capital and labor costs required to maintain the experimental facilities. Furthermore, when batch processing is implemented to increase efficiency, it is difficult to insert urgent samples.

[0003] In recent years, StoA (Sample-to-Answer) systems, which perform all processes from sample introduction to measurement and data acquisition in a fully automated manner, have begun to appear in a variety of fields. StoA systems sometimes use a flow channel chip, which integrates chambers, flow channels, and reagents. The introduction of a flow channel chip offers the following advantages: (i) measurements can be easily performed even by non-experts; (ii) data can be acquired in a short time; (iii) highly portable devices can be designed; (iv) variability due to the procedure is reduced; and (v) reagent storage is simplified.

[0004] Potential applications of StoA flow channel chips include forensic science, in vitro diagnostics, identification of plant and animal species, biodefense, medicine, biotechnology, life sciences, defense, public health, and agriculture. When performing genetic analysis using StoA flow channel chips, it is desirable to dispose of part or all of the flow channel that comes into direct contact with the sample after each measurement to prevent sample contamination between analyses. Measures to reduce the cost of disposable chips include designs that are easy to manufacture and the use of inexpensive materials.

[0005] However, in such inexpensive chips, the pressure resistance of the chip is limited by factors such as valves and chip bonding strength. For example, in Patent Document 1, the channel chip has a simple structure consisting of two sheets of thermoplastic resin bonded together. The pressure resistance of this chip is determined by the valve and is described as 68 kPa. Furthermore, in Patent Document 2, for example, the pressure resistance of the channel chip is described as, for example, 124 kPa. In contrast, in large-scale channel systems, such as liquid chromatography, the pressure that can be used to transport solutions exceeds several MPa. Furthermore, in the case of spin columns, which are widely used in nucleic acid purification, a pressure of up to 500 kPa can be applied. Thus, the pressure that can be used when performing sample processing on a StoA channel chip tends to be lower than that on a benchtop.

[0006] In StoA systems, the transport of a solution must be completed automatically within a limited space and pressure, so the presence of air bubbles in the flow path is undesirable. This is because there is a concern that air trapped in the flow path may cause incomplete transport of the solution or unexpected behavior. To solve the problems caused by the presence of air in the flow path, according to Non-Patent Document 1, a material that allows air to escape is used in the flow path. Furthermore, according to Non-Patent Document 2 and Patent Document 3, a structure for evacuating air is installed in the flow path. It is also possible to solve this problem by applying high pressure to compress or move the air.

[0007] U.S. Patent No. 10,233,491 U.S. Patent No. 9,354,199 Japanese Patent No. 6,613,212

[0008] “PDMS membranes with tunable gas permeability for microfluidic applications - RSC Advances (RSC Publishing) DOI:10.1039 / C4RA1293B,” RSC Adv., 2014, 4, 61415“Integrated Microfluidic System for Rapid Forensic DNA Analysis: Sample Collection to DNA Profile / Analytical Chemistry (acs.org),”Anal. Chem. 2010, 82, 16, 6991-6999

[0009] In the purification and recovery of nucleic acids using a purification membrane such as silica, a high recovery efficiency can be achieved with a simple channel structure. However, when this purification method is implemented in a channel with limited space for air to escape, the following problem occurs. If air enters the chamber containing the membrane, the air must pass through the membrane chamber. For example, when air passes through the membrane while it is wet with the lysate after the lysate has passed through the membrane, the pressure applied is the Laplace pressure P defined by the following equation (1): Ｌ It is necessary to exceed this.

[0010]

[0011] Here, θ, d, and γ are the contact angle between the liquid wetting the membrane and the membrane, the pore size of the membrane, and the surface tension of the liquid, respectively. When a fine silica membrane is used for purification, d is small, so it is thought that the Laplace pressure will be significantly high.

[0012] In this regard, as in the techniques disclosed in Patent Document 3, Non-Patent Document 1, and Non-Patent Document 2, by providing a structure for removing air, it is possible to prevent air from passing through the membrane.

[0013] However, this approach requires the introduction of additional structures into the flow channels, which may limit the materials that can be used for these structures, and may increase the cost and size of the chip.

[0014] Furthermore, in the case of Patent Document 1, the purification membrane installed in the purification chamber does not cover the entire flow path. In this case, the air passes over the side of the purification membrane, so the above-mentioned problem does not occur, but the proportion of dissolved product that comes into contact with the membrane decreases, which raises concerns about a decrease in recovery rate. In light of this situation, the present disclosure proposes a technology that prevents fluids (air, nitrogen, and other gases) from passing over the purification membrane even without an additional flow path structure.

[0015] In order to solve the above problems, the present disclosure proposes, as an example, a method for controlling liquid transport in a flow path of a biomolecule analyzer using a computer, wherein the biomolecule analyzer has a first chamber that contains a first liquid, a second chamber that contains a second liquid, a membrane chamber having a purification membrane, and a waste liquid chamber, and the method includes: controlling, by a computer, the transport of the second liquid at least past the confluence of a first flow path connecting the first chamber to the waste liquid chamber and a second flow path extending from the second chamber; discharging a fluid different from the first and second liquids from the second flow path; controlling, by a computer, the transport of the first liquid from the first chamber to the waste liquid chamber via the membrane chamber; and controlling, by a computer, the transport of the second liquid in the second chamber from the second chamber to the membrane chamber.

[0016] Further features related to the present disclosure will become apparent from the description of this specification and the accompanying drawings. Also, aspects of the present disclosure are achieved and realized by the elements and combinations of various elements and the aspects of the following detailed description and the appended claims. The description of this specification is merely exemplary and does not limit the scope or application of the claims of the present disclosure in any way.

[0017] According to the technology of the present disclosure, air can be prevented from crossing the purification membrane without the need for an additional flow path structure.

[0018] 1 is a diagram showing an example of the configuration of a biomolecule analyzer 100 according to the present embodiment; FIG. 2 is a diagram showing a portion of the biomolecule analyzer 100 and a derivative of a channel chip 114; FIG. 3 is a diagram showing an example of a procedure for performing bioanalysis using the biomolecule analyzer 100; FIG. 4 is a diagram showing an example of the internal configuration of a computer 115; FIG. 5 is a diagram showing an example of the configuration of a purification system 301 including the main parts of the channel chip 114 according to Example 1, the main parts of which are the channel chip 114 at the completion or during the sample dissolving procedure 202; FIG. 6 is a diagram showing purification processes I to V according to Example 1; FIG. 7 is a flowchart corresponding to the process shown in FIG. 6; FIG. 8 is a diagram for explaining a purification process according to a comparative example; FIG. 9 is a flowchart of the purification process according to FIG. 8; FIG. 10 is a schematic diagram showing the position of a solution during transport and the corresponding pressure changes measured in the experiment of Example 1; FIG. 11 is a diagram showing an example of the configuration of a purification system 400 including the main parts of the channel chip 114 according to Example 2, the main parts of which are the channel chip 114 at the completion or during the sample dissolving procedure 202; FIG. 12 is a diagram for explaining purification processes I to V according to Example 2. 14A and 14B are diagrams illustrating an example of the configuration of a purification system 500 including a main part of a channel chip 114 according to Example 3, the main part being the channel chip 114 at the completion or during the sample dissolving procedure 202. FIG. 15A is a diagram illustrating purification processes I to VI according to Example 3. FIG. 15B is a flowchart corresponding to the purification process shown in FIG. 14A. FIG. 15C is a diagram illustrating an example of the configuration of a purification system 600 including a main part of a channel chip 114 according to Example 4, the main part being the channel chip 114 at the completion or during the sample dissolving procedure 202. FIG. 15D is a diagram illustrating purification processes I to V according to Example 4. FIG. 15E is a diagram illustrating purification processes I to V according to Example 5. FIG. 15F is a diagram illustrating purification processes I to V according to Example 6.

[0019] This embodiment proposes, through each example, a technique for installing a purification membrane on a channel chip by reducing the pressure required for liquid transport in the channel chip, eliminating the need for structural robustness in the channel chip. First, the features of the biomolecular analysis device according to this embodiment will be described, followed by a description of each example. In the accompanying drawings, functionally identical elements may be designated by the same number. While the accompanying drawings show specific embodiments and implementation examples in accordance with the principles of this disclosure, these are intended to facilitate understanding of this disclosure and are not intended to limit the scope of this disclosure. Furthermore, while this embodiment is described in sufficient detail to enable those skilled in the art to implement this disclosure, it should be understood that other implementations and forms are possible, and that configuration and structure changes and various element substitutions are possible without departing from the scope and spirit of the technical concept of this disclosure. Therefore, the following description should not be interpreted as being limited to these examples.

[0020] Furthermore, in embodiments of the present disclosure, computer-controlled operations may be implemented in software running on a general-purpose computer, in dedicated hardware, or in a combination of software and hardware.

[0021] (1) Features of the Biomolecule Analysis Device <Channel Chip> In this embodiment, a "channel chip (or simply chip)" refers to a disposable or multiple-use cartridge that contains reagents, chambers, and channels. The channel chip may contain a power source for transporting the solution. Some or all of the reagents may be present within the chip. Some of the chambers may be equipped with temperature control functions, molecular capture functions, detection functions, and voltage application functions.

[0022] The material of the channel chip is not particularly limited as long as it is a material commonly used in the relevant technical field. For example, materials that have low DNA adsorption, such as polypropylene, cyclic olefin polymer (COP), cyclic olefin copolymer (COC), polycarbonate, polyethylene terephthalate, and polyurethane, are preferably used. It is also desirable to suppress adsorption by modifying the surface so that it is negatively charged. Other materials include, for example, metals such as gold, silver, copper, aluminum, tungsten, molybdenum, chromium, platinum, titanium, and nickel; alloys such as stainless steel, Hastelloy, Inconel, Monel, and duralumin; silicon; glass materials such as glass, quartz glass, fused silica, synthetic quartz, alumina, sapphire, ceramics, forsterite, and photosensitive glass; plastics such as polyester resin, polystyrene, polyethylene resin, ABS resin (Acrylonitrile Butadiene Styrene resin), dimethylpolysiloxane (PDMS), nylon, acrylic resin, fluororesin, polycarbonate resin, polyurethane resin, methylpentene resin, phenolic resin, melamine resin, epoxy resin, and vinyl chloride resin; agarose, dextran, cellulose, polyvinyl alcohol, nitrocellulose, chitin, chitosan, or any combination thereof.

[0023] If the cross-sectional area of ​​the flow channel is too large, there is a concern that residual liquid or sample loss may occur. Also, if the cross-sectional area is too large, it becomes difficult to apply pressure. On the other hand, if the cross-sectional area is too small, there are problems such as high pressure being required to transport the solution or the time required for transport being long. Therefore, the cross-sectional area of ​​the flow channel is set to 1 μm or less. ２ ~314mm ２ In a more preferred configuration, the cross-sectional area is 400 μm ２ ~100mm ２ , and more preferably, the cross-sectional area is 0.01 mm ２ ~10mm ２ The range can be:

[0024] If the length of the flow channel is too short, it is difficult to incorporate elements such as valves, and if it is too long, there is a concern that the chip will become too large. Therefore, the length of the flow channel connecting the components of the chip can be set to 1 μm to 100 cm. A more preferable range is 1 mm to 50 cm. An even more preferable range is 5 mm to 30 cm.

[0025] <Chamber> A chamber is a space capable of storing liquids or solids, and has a diameter equal to or larger than that of a flow channel. Reagents may be stored in the chamber, and PCR, lysis, purification, etc. may be performed in the chamber.

[0026] The capacity of the chamber is, for example, 0.01 μL to 10 L. If the capacity is greater than 10 L, the device becomes difficult to carry. In a more preferred example, the capacity of the chamber can be 0.1 μL to 2 L. Note that it is not necessary for all chambers to be installed within the chip, and they may be provided in the device body or in a separate, independent chip.

[0027] When a chamber is installed inside a chip, the capacity of the chamber can be 0.01 μL to 50 mL. If the capacity is greater than 50 mL, the chip becomes too large and difficult to store.

[0028] The chip contains one or more reagents stored in one or more reagent storage chambers. Because unintended mixing of these reagents can lead to performance degradation or other unexpected results, it is desirable for them to be separated by a barrier mechanism consisting of a valve, film, air, or a channel narrow enough to prevent spontaneous mixing, or a combination of these, until just before use. Furthermore, isolating the reagents from the outside air allows for long-term storage and chip portability.

[0029] Similarly, when reagents are stored outside the chip (when the chambers are external to the flow path chip), it is desirable that the reagents be stored in isolation from the outside air, separated from other purification system components by valves, films, air, etc.

[0030] <Purification System> In this embodiment, the "purification system" refers to a system that captures biomolecules contained in a lysate stored in a lysis chamber using a purification membrane provided on a channel chip and a membrane chamber that houses the purification membrane, and then washes the lysate with a washing solution stored in a washing solution chamber. The lysis chamber and the washing solution chamber may be provided outside the channel chip, or either one may be provided inside the chip.

[0031] The purification system may be configured to selectively recover specific biomolecules. For example, DNA can be selectively extracted from a liquid containing proteins, DNA, ions, etc. The purification system may also be a purification system for recovering RNA, biopolymers (nucleic acids, proteins, lipids, polysaccharides), or molecules containing biomonomers (amino acids, lipids, sugars, nucleic acid bases) and their derivatives in their structure. It may also be a system that recovers multiple types of these molecules. One example is a system that uses a silica membrane as the purification membrane and can recover DNA and RNA.

[0032] <Sample Type> The sample provided to the purification system according to this embodiment is not particularly limited as long as it is a biological sample. The biological source from which the sample is derived is also not particularly limited, and samples derived from any biological organism, such as vertebrates (e.g., mammals, birds, reptiles, fish, amphibians, etc.), invertebrates (e.g., insects, nematodes, crustaceans, etc.), plants, protozoa, fungi, bacteria, and viruses, can be used. When collecting a sample, a swab, filter paper, cloth, or the like can be used as a carrier, and the carrier may be introduced into the purification system.

[0033] <Lysate> When a sample is delivered to the membrane chamber, it must be in a form that allows it to flow through the flow path. Therefore, if the sample is solid (e.g., a swab sample), it is preferable to dissolve or suspend the solid sample in a lysis buffer to form a fluid lysate. The sample does not need to be completely dissolved; any solid or highly viscous portions may be retained in the lysis chamber after lysis. Furthermore, if the sample is a gaseous sample (e.g., air, exhaled breath, etc.), it is preferable to suspend cells contained in the gaseous sample in a solvent to form a liquid sample. Preparation methods for converting a sample into a lysate are routine in the art and readily understood by those skilled in the art. For example, the lysis buffer may contain a chlorinated material such as calcium hypochlorite. As another example, the substance may contain enzyme activity such as DNAase, RNAase, or protease. If necessary, the lysis buffer may contain substances that facilitate the release of biomolecules, such as chaotrope, surfactants, or KOH, or substances that facilitate the binding of nucleic acids to the purification membrane. If necessary, the mixture may be subjected to treatment such as heating and stirring.

[0034] In this embodiment, the term "lysate" refers to a substance obtained by converting a biological sample into a liquid with a viscosity of 100,000 mPa·s or less using a lysis buffer. More preferably, the lysate may have a viscosity of 10,000 mPa·s or less. Even more preferably, the lysate may have a viscosity of 1,000 mPa·s or less.

[0035] <Solution Transport Control> A valve may be used to control the transport of the solution. Alternatively, the solution may be controlled by flow path resistance. In this embodiment (each example), air pressure is used as the power for transporting the solution. Alternatively, air pressure, mechanical compression, centrifugal force, or the like may be used as the power for transporting the solution.

[0036] If the pressure used for transport is too high, it will exceed the chip's pressure resistance, while if it is too low, the time required for transport will be long, leading to extended measurement times. Therefore, the pressure used for transport can be in the range of 0.1 kPa to 1 MPa. Solution transport can be completed at a narrower range of 0.1 kPa to 500 kPa, or an even narrower range of 0.1 kPa to 200 kPa. Furthermore, the time required for transport can be up to 1 hour per step, 10 minutes in a narrower range, or within 5 minutes in an even narrower range. Furthermore, if the volume of liquid transported is too large, the time and pressure required for transport will increase, leading to higher chip costs and reagent costs. Therefore, the volume of liquid transported per reagent can be within 1 L, within a narrower range of 10 mL, or within an even narrower range of 2 mL.

[0037] <Cleaning Solution> In this embodiment, the term "cleaning solution" refers to a liquid used to wash away substances that adhere to the purification membrane and are not required for subsequent processes. Note that the cleaning solution does not necessarily have to be able to wash away all of the unnecessary substances, and may instead wash away some or all of the necessary substances.

[0038] According to the above formula (1), in this embodiment, a cleaning liquid having the following characteristics can be used. First, a liquid that evaporates faster than the dissolved product is preferable. Also, it is desirable that the dissolved product and the cleaning liquid are compatible. Furthermore, it is desirable that the contact angle with the purified membrane is smaller (low wettability) than the dissolved product, or that the interfacial tension is low. Examples of cleaning liquids that satisfy these requirements include ethanol and isopropanol. Furthermore, liquids containing 10% or more of these alcohols can also be used as cleaning liquids. However, solutions that do not satisfy the above conditions may also be used.

[0039] The number of cleaning liquids may be one or two or more. Using two or more cleaning liquids enables more efficient cleaning. They may be stored in one chamber or in two or more chambers. When two or more cleaning liquids are used, air may or may not be trapped between them. However, if the second or subsequent cleaning liquids have a faster evaporation rate, a lower surface tension, or a smaller contact angle with the purification membrane than the first cleaning liquid, it is desirable to transport them continuously.

[0040] <Purification membrane> One type of membrane can be a silica membrane. Other examples of purification membranes include solid substrates whose main component is cellulose, capable of adsorbing DNA, carboxylated particles, and ion exchange resins. In particular, membranes having hydroxyl groups or silica groups on their surfaces can be used. Any membrane can be used as long as it can retain particles of 100 μm or more. It is also preferable that the thickness is 1 μm or more. Furthermore, since the finer the mesh, the more efficiently DNA can be recovered, a membrane that can retain particles of 10 μm or more, more preferably 1 μm or more, and even more preferably 0.1 μm or more can be used.

[0041] If the volume of the purification membrane is too small, the amount of biomolecules that can be adsorbed will be small. On the other hand, if the volume is too large, there is a concern that unintended molecular adsorption may occur during purification or subsequent steps, and that the efficiency of solution transport may be reduced. In each example described below, an area of ​​12.5 mm ２ However, for example, a 1 mm ２ ~314mm ２ The membrane may be any membrane, and there is no limitation on the size.

[0042] <Detection Method> PCR amplification is performed downstream of this purification system. After amplification, detection is performed by capillary electrophoresis (CE). Other methods that can be used include massively parallel sequencing (MPS), pyrosequencing, Sanger sequencing, nanopore sequencing, chromatography, electrical measurements, spectroscopy, NMR, and restriction fragment length polymorphisms (RFLP).

[0043] <Other Supplementary Notes> In all drawings illustrating this embodiment, components having the same function are assigned the same reference numerals, and repeated explanations are omitted as much as possible. Below, each example of this embodiment will be described in detail with reference to the accompanying drawings. The measurement methods, device structures, types of substances, and materials described in each example are examples for embodying the concept of this embodiment, and do not strictly specify the measurement principles, device materials, dimensions, etc. Furthermore, the specific pressure values ​​described in each example are examples for embodying the concept of this embodiment, and do not strictly specify them. Furthermore, the specific sample types, purification kit compositions, and liquid volumes described in each example are examples for embodying the concept of this embodiment, and are not examples for strictly defining the chemical composition or time. Furthermore, the specific types of measurement targets, types of solutions, and their concentrations described in each example are examples for embodying the concept of this embodiment, and are not examples for strictly defining the chemical composition.

[0044] 1 is a diagram showing an example of the configuration of a biomolecule analyzer 100 according to this embodiment. The biomolecule analyzer 100 includes a channel chip 114 and a computer 115 for performing biomolecule analysis.

[0045] The flow channel chip 114 has a lysis chamber 101 for introducing and lysing a collected sample, a membrane chamber 108 containing a purification membrane 102, a reaction chamber 103 for performing DNA amplification and the like, a waste chamber 107, and a port 109 fluidly connected to the outside of the chip. Note that the arrangement of each chamber in the flow channel chip 114 and the flow channel connections between the chambers differ depending on the embodiment, and therefore the chamber arrangement, flow channel connections, and the transport operation of the lysate and washing solution will be described later.

[0046] A glass fiber membrane GF / F sold by Whatman, cut to a diameter of 4 mm, was installed as the purification membrane 102. Other membranes may also be used for the purification membrane 102. The diameter of the purification membrane (the longitudinal length in the case of a rectangular membrane) may be in the range of 0.1 mm to 100 mm.

[0047] Solution transport is achieved by applying pressure via port 109, allowing reagents, amplification products, and the like to be exchanged with the outside of the chip. Pressure can be applied using a pressure generator located outside the chip. The channel chip 114 shown in FIG. 1 incorporates chambers 104, 105, and 106 for storing reagents and the like. Regarding the membrane chamber 108, the membrane-separated space near the lysis chamber 101 has a capacity of 10 μL, while the membrane-separated space near the waste chamber 107 has a capacity of 10 μL. The lysis buffer chamber 104 stores a lysis buffer 110 for lysing the sample, the wash solution chamber 105 stores a wash solution 111, and the reagent chamber 106 stores an eluate or a reagent 112 used in the reaction. Some of the above functions may be integrated into the same chamber. The positional relationship of the above-described chip components is not limited to that shown in FIG. 1 , and the channel connections may differ from those shown in FIG. 1 .

[0048] 2 is a diagram showing a portion of the biomolecule analyzer 100 and a derivative form of the channel chip 114. In FIG. 2, the lysis buffer chamber 104, the washing solution chamber 105, and the waste fluid chamber 107 are provided outside the channel chip 114. By adopting such a configuration, the advantages of miniaturization and cost reduction of the channel chip 114 can be enjoyed.

[0049] <Example of Bioanalysis Procedure> Fig. 3 is a diagram showing an example of a procedure for performing bioanalysis using the biomolecule analysis device 100. As shown in Fig. 3, the bioanalysis includes, for example, a sample reception procedure 201, a sample dissolution procedure 202, a sample preparation procedure 203, a sample amplification procedure 204, and a sample detection procedure 205.

[0050] In the sample receiving procedure 201, before or after the sample is stored in the lysis chamber 101, the lysis buffer 110 is transported from the chamber 104 to the lysis chamber 101 (a step of introducing the sample into the lysis chamber 101).

[0051] Next, lysis begins in the sample lysis step 202. In the sample preparation step 203, the lysate 113 is transferred from the lysis chamber 101 to the purification membrane 102, where the DNA binds to the membrane and is purified. After purification, a step of drying the wash solution or the like may be included. The eluted DNA is transported to the reaction chamber 103.

[0052] In sample amplification step 204, the purified DNA is amplified. In sample detection step 205, the amplified DNA is measured. Sample detection step 205 may be performed within the channel chip 114, or may be transported to a detection unit outside the channel chip 114 and performed in a separately provided measurement unit. Steps 201 to 205 may be performed in parallel. Also, some steps may be omitted, or other steps may be incorporated.

[0053] 4 is a diagram showing an example of the internal configuration of the computer 115. The computer 115 includes a processor (not shown), a user interface 1151, and a database (storage device) 1152.

[0054] The user interface 1151 has an input screen and an output screen, and can accept parameters related to the implementation procedure in Fig. 3 from the user, such as the time, temperature, pressure, flow rate, and procedure of each step, and store them in a database 1152. In addition, various parameters can be saved in advance in the database 1152.

[0055] The computer 115 can open and close the valves of the channel chip 114, control the temperature, and control the applied pressure and flow rate based on various parameters recorded in the database 1152. The computer 115 can also automatically control all of the implementation procedures shown in Fig. 3. Note that a user may assist and implement some of the implementation procedures shown in Fig. 3.

[0056] (2) Example

[0057] The first embodiment will be described with reference to FIGS.

[0058] <Example of configuration of purification system 301 including main parts of flow path chip 114> Figure 5 is a diagram showing an example of configuration of purification system 301 including main parts of flow path chip 114 according to Example 1, when sample dissolution procedure 202 is completed or is being performed.

[0059] The purification system 301 includes a lysis chamber 101, a washing solution chamber 105, a waste solution chamber 107, a membrane chamber 108, valves 302, 303, 304, 305, and 315, flow paths 306, 307, 308, 309, 310, 311, and 312 connecting the chambers, and a port 109. The number of port 109 does not have to be one, and one port may be provided for each function. For example, the flow paths 311 and 310 may each be provided with a dedicated port.

[0060] The lysing chamber 101 holds a lysate 113 therein, and the washing liquid chamber 105 holds a washing liquid 111 therein. A liquid level detection sensor 313 is installed in the waste liquid chamber 107, and a liquid level detection sensor 314 is installed in the lysing chamber 101. One or more liquid level detection sensors may be installed, or none may be installed. A liquid level detection sensor may also be installed in a chamber other than the lysing chamber 101 or the waste liquid chamber 107.

[0061] As shown in Figure 5, a channel 306 extending from the outlet of the lysis chamber 101 and a channel 307 extending from the outlet of the washing solution chamber 105 join at a connection point (junction) 316 to form a channel 308, which is connected to the inlet of the membrane chamber 108. A channel 309 extending from the outlet of the membrane chamber 108 connects to an inlet at the bottom (bottom surface: not necessarily the bottom) of the waste chamber 107. Two channels 310 and 311 extend from the port 109, with the channel 310 connecting to the top (top surface: not necessarily the top surface) of the waste chamber 107 and the channel 311 connecting to the inlet of the washing solution chamber 105. Furthermore, the channel 312 connects to the top (top surface: not necessarily the top surface) of the waste chamber 107.

[0062] As shown in FIG. 5, valve 302 is provided in flow path 307, valve 303 in flow path 311, valve 304 in flow path 310, valve 305 in flow path 306, and valve 315 in flow path 312, respectively.

[0063] <Purification Process> Fig. 6 is a diagram showing purification processes I to V according to Example 1. Fig. 7 is a flowchart corresponding to the processes shown in Fig. 6. Note that the opening and closing of each valve in each process and the transport control of the lysate 113 and the washing solution 111 by applying pressure are performed by the computer (processor) 115.

[0064] (i) Process I Process I shows the state during or immediately after the completion of dissolution in the sample dissolution procedure 202. At this time, valves 302, 303, 304, 305, and 315 are closed (step 701).

[0065] (ii) Process II In Process II, valves 302, 303, and 315 are opened (Step 702), and wash solution 111 is pumped from wash solution chamber 105 to membrane chamber 108 at a pressure of 30 kPa or less for 30 seconds or less (Step 703). At this time, air in flow channel 307 moves to flow channel 308 or 306, and flow channel 307 is filled with wash solution 111. In Process II, it is important to evacuate air from flow channel 307 in order to achieve continuous transport without trapping air between lysate 113 and wash solution 111.

[0066] (iii) Process III In Process III, valves 302, 303, and 315 are closed, and valves 305 and 304 are opened (Step 704), and 900 μL of lysate 113 is transported from lysis chamber 101 to waste chamber 107 via membrane chamber 108 at an applied pressure of −60 kPa for 1 minute (Step 705). The amount of lysate 113 transported (900 μL) is controlled by computer 115 based on the liquid level detection sensor 314 and the application time (1 minute) of a predetermined pressure (60 kPa).

[0067] (iv) Process IV In Process IV, before the air following the lysate 113 passes the branch point (junction point 316) of flow paths 306 and 308 and flow path 307, valves 304 and 305 are closed, and valves 302, 303, and 315 are opened (step 706). 500 μL of washing solution 111 is transported from washing solution chamber 105 to waste chamber 107 via membrane chamber 108 at an applied pressure of 60 kPa for 3 minutes (step 707). The amount of washing solution 111 transported (500 μL) is controlled by computer 115 based on the liquid level detection sensor 314 and the application time (3 minutes) of a predetermined pressure (60 kPa). By performing the operations shown in Processes I to IV, continuous transport can be achieved without trapping air between the lysate 113 and washing solution 111.

[0068] <Comparative Example> Fig. 8 is a diagram for explaining a refining process according to a comparative example, and Fig. 9 is a flowchart of the refining process according to Fig. 8 .

[0069] (i) Process I Process I shows the state during or immediately after the completion of dissolution in the sample dissolution procedure 202 (step 901). At this time, valves 302, 303, 304, and 305 are closed.

[0070] (ii) Process II In process II, valves 304 and 305 are opened (step 902), and lysate 113 is transported from lysis chamber 101 to waste chamber 107 via membrane chamber 108 (step 903).

[0071] (iii) Process III In process III, once the transfer of lysate 113 is complete, valves 305 and 304 are closed (step 904).

[0072] (iv) Process IV In process IV, valves 302, 303, and 315 are opened (step 904), and the cleaning solution 111 is transported from the cleaning solution chamber 105 to the waste chamber 107 via the membrane chamber 108 (step 905).

[0073] (v) Process V Process V shows the state after the entire amount or a predetermined amount of the cleaning solution 111 has been transported to the waste chamber 107 via the membrane chamber 108 .

[0074] In the comparative purification process, air present in channels 307 and 308 enters between the lysate 113 and the cleaning solution 111. Therefore, in order for the cleaning solution 111 to reach the membrane, it is necessary to overcome the Laplace pressure generated between the lysate 113 and the purification membrane 102.

[0075] <Regarding Technical Effects> In order to confirm the technical effects of the transport operation of the lysate and washing solution according to Example 1, a comparative experiment was conducted using a simple flow path system to compare the pressure required to transport the solution when air was introduced into the flow path and when it was not. Note that while the technical effects will be discussed here based on the experimental results of Example 1, the technical effects listed here also apply to Examples 2 to 4 and derivative examples described below.

[0076] (i) Experimental conditions The flow path system used the lysate prepared according to the Qiagen QIK (Qiaamp Investigator Kit DNA) protocol and the QIK cleaning solution (AW1). The lysate and cleaning solution were each placed in a 1.5 mL tube, pressurized with a syringe pump, and transported to a polycarbonate membrane chamber. The membrane chamber can hold approximately 20 μL of solution, and is separated in the center by a membrane, with both the upstream and downstream volumes being approximately 10 μL. The liquid contact area of ​​the purification membrane is 3.1 mm ２ The lysate and washing solution that passed through the membrane chamber were collected in a waste tube installed behind the membrane chamber. The extrusion speed of the syringe pump was set to 4 mL / min.

[0077] In a comparative example (Procedure A) different from that of Example 1, 100 μL of lysate, 100 μL of air, and 500 μL of cleaning solution (QIK Cleaning Solution AW1) were sequentially delivered to the membrane chamber, and the change in pressure over time was measured. To replicate the delivery method of Example 1, Procedure B involved consecutive delivery of 100 μL of lysate and 500 μL of cleaning solution without air between them. Whatman's glass fiber membrane GF / F was used as the membrane.

[0078] (ii) Experimental Results Figure 10 is a diagram showing the position of the solution during transport and the corresponding measured pressure changes in the above-mentioned experiment. In Figure 10, the dashed line graph shows the results of Procedure A (Comparative Example), and the solid line graph shows the results of Procedure B (Example 1).

[0079] Because the extrusion speed of the syringe pump was greater than the movement speed of the solution, the pressure increased over time while the lysate passed through the membrane, reaching 20 kPa just before the lysate was completely removed from the membrane (Figure 10 (solid line / dashed line), step (a) → (b)).

[0080] In the case of Procedure A, once the dissolved product had completely escaped and air had entered the membrane, the liquid surface stopped moving. Further, as the syringe pump continued to be pressed, air slowly began to cross the membrane at pressures above 80 kPa. When the pressure reached 180 kPa, the washing solution reached the membrane chamber (Figure 10 (dashed line), steps (c) → (d)). After the washing solution had completely escaped across the membrane, the pressure was confirmed to drop sharply to around 30 kPa (Figure 10 (dashed line), step (d)).

[0081] In the case of Procedure B, when the dissolved product is completely removed and the washing solution enters the membrane, the solution does not stop (Figure 10 (solid line) step (b) → (c)). The pressure did not exceed 40 kPa until the washing solution was completely removed. Separately, the pressure required to transport the washing solution alone was confirmed, and the transport was completed within 40 kPa (data not shown).

[0082] (iii) Discussion of Experimental Results As described above, from the experimental results of Procedure A (Comparative Example) and Procedure B (Example 1), it was confirmed that the required pressure can be reduced to one-third by using the conveying method according to Example 1 (as well as other examples described below).

[0083] The reason why high pressure is not required to allow air to enter after the cleaning solution has been removed is that the evaporation rate of the cleaning solution is much higher than that of the dissolved product. The cleaning solution (QIK, AW1) is composed of more than 50% ethanol by volume.

[0084] In this example, the lysate and washing solution were transported continuously, reducing the maximum pressure required to transport the two types of liquid. The effect of this example is not limited to the kit described above, and the composition of the lysate or washing solution may be different. When the second liquid (washing solution) is completely removed before the sample detection step 205, the following requirements can be met for a combination of the first liquid (lysate) and the second liquid that is more likely to achieve a pressure reduction effect: (a) The evaporation rate of the second liquid is faster than that of the first liquid; (b) The surface tension of the second liquid is lower than that of the first liquid; (c) The contact angle of the second liquid with respect to the membrane is larger than that of the first liquid; (d) The viscosity of the second liquid is lower than that of the first liquid.

[0085] When the main component of the first liquid is water, an example of a second liquid that satisfies one or more of the above-mentioned conditions is a solution containing 10% or more of an alcohol having 4 or less carbon atoms. More preferably, an example of a solution containing 30% or more of an alcohol having 3 or less carbon atoms. Even more preferably, an example of a solution containing 40% or more of ethanol. Alternatively, an appropriate second liquid may be selected depending on the composition of the first liquid, or the first liquid may be selected depending on the composition of the second liquid.

[0086] According to Example 1 (experimental results) of the present disclosure, the required pressure can be reduced. Furthermore, by reducing the required pressure, it becomes possible to install a high-density purification membrane on a structurally weak channel chip. Here, examples of structurally weak chips include chips made by bonding valves and chips together or using soft materials. Manufacturing structurally weak chips has advantages such as reduced chip costs and higher chip functionality. Furthermore, by reducing the required pressure, chemical reactions that are inhibited by applying high pressure can be efficiently promoted. Furthermore, applying high pressure can facilitate the measurement of biomolecules.

[0087] In addition to reducing the pressure, the technology disclosed herein can also shorten the time it takes to transport the solution, and by using the technology disclosed herein, it is possible to prevent the membrane from drying out after the lysate has passed through it and before washing, thereby providing benefits such as increased yield of biomolecules and improved washing efficiency.

[0088] (iv) Membrane for DNA Recovery In the biomolecule analyzer 100, highly efficient DNA recovery can be achieved by installing a high-density membrane. "Particle retention size" is an index of mesh size. This means that particles with a diameter equal to or larger than the value indicated in the particle retention size are retained by the membrane. In other words, the smaller this value, the finer the membrane. The membrane used as a purification membrane can have a particle retention size of 100 μm or more. For example, it is desirable to use a membrane with a mesh size finer than the "Fusion 5" membrane sold by Whatman. Another membrane capable of more efficient DNA recovery is the "GF / D" membrane sold by Whatman. The particle retention size of this membrane is 2.7 μm. Another membrane capable of more efficient DNA recovery is the "GF / F" membrane sold by Whatman. The particle retention size of this membrane is 0.7 μm.

[0089] Furthermore, highly efficient DNA recovery can be achieved by stacking multiple membranes. For example, in the case of GF / F, stacking two or more membranes allows for more efficient DNA recovery than purification using only one membrane. The membrane thickness is preferably 0.001 mm or more. In the case of GF / F, the membrane thickness is 0.6 mm.

[0090] (v) Regarding the membrane chamber 108: The volume of the membrane chamber 108 is 30% or less of the volume of the lysis chamber 101 or the wash solution chamber 105. The cross-sectional area of ​​the channel through which the two solutions pass in common, in the direction perpendicular to the flow direction, is 50% or less of the maximum cross-sectional area of ​​the chamber, and the volume of the channel through which the two solutions pass in common is 50% or less for each of the two solutions. Therefore, the lysate and wash solution are not mixed by more than 50% before reaching the waste chamber. Therefore, transporting the solution using the technology disclosed herein does not significantly reduce the DNA binding efficiency.

[0091] The size of the membrane chamber 108 is 0.1 μL to 1 mL. The smaller the membrane chamber 108, the less loss occurs during elution, so the size of the membrane chamber 108 is more preferably 0.1 μL to 100 μL, and even more preferably 0.1 μL to 30 μL.

[0092] (vi) Regarding the Flow Channel The volume of the flow channel 307 is 0.1 μL to 1 L (the volume of the flow channel will be larger if a long flow channel is used). If the volume of the flow channel 307 is larger than 1 L, it will take a long time to transport the solution. Conversely, if the volume is small, it will be necessary to narrow the flow channel width or reduce the distance between the chambers, reducing the design freedom. Furthermore, if a chamber exists outside the chip, a large volume is desirable. If the flow channel is too narrow, there is a risk of unexpected behavior, such as clogging of the flow channel. A more preferable flow channel volume is 1 μL to 100 mL, and in another embodiment, 30 μL to 100 mL. Furthermore, the volume of the flow channel 307 can be the same as or larger than the volume of the space 108_1 above the purification membrane 102 in the membrane chamber 108.

[0093] In addition, when drying is performed after purification, it is desirable to ensure a certain degree of width of the flow channels. In this case, the width of the flow channels 302 and 303 is 0.03 mm. ２ It is desirable that:

[0094] <Regarding Process Switching Control> (i) Control Using a Liquid Level Detection Sensor A liquid level detection sensor can be used to switch between processes I, II, III, IV, and V in Fig. 6. For example, when switching from process III to IV, liquid level detection sensors 313 and 314 are used. Sensors that detect electrical, optical, acoustic, or mechanical levels can be used as the liquid level detection sensors 313 and 314.

[0095] When the liquid level detection sensor 313 is used, a signal generated when the liquid level in the waste chamber 107 exceeds the liquid level detection sensor 313 can be used when switching from process III to IV. However, the liquid level detection sensor 313 needs to be installed at a position where it can detect the liquid level in the waste chamber 107 when the remaining amount of the lysate 113 in the lysis chamber 101 falls below 10%.

[0096] Furthermore, when the liquid level detection sensor 314 is used, a signal indicating that the liquid level in the dissolution chamber has fallen below the liquid level detection sensor 314 can be used when switching from process procedure II to III. However, the liquid level detection sensor 314 needs to be installed in a position where it can detect the liquid level when the remaining amount of the lysed product 113 in the dissolution chamber 101 falls below 10%.

[0097] (ii) Control by Pressure, Time, etc. Preset pressure and time may be used to switch between Processes I, II, III, and IV in Figure 6. The pressure and time may also be adjustable by an operator. Furthermore, the dissolution chamber 101 may be equipped with a mechanism for measuring viscosity. Furthermore, flow rate sensors and pressure sensors may be installed at specific locations to define the timing of process switching.

[0098] A second embodiment will be described with reference to FIGS. 7, 11, and 12. FIG.

[0099] <Example of configuration of purification system 301 including main parts of flow path chip 114> Figure 11 is a diagram showing an example of configuration of a purification system 400 including main parts of the flow path chip 114 according to Example 2, which are the main parts of the flow path chip 114 when the sample dissolution procedure 202 is completed or is being performed.

[0100] The purification system 400 includes a lysis chamber 101, a washing solution chamber 105, a waste solution chamber 107, a membrane chamber 108, valves 401, 402, 403, 404, 405, and 406, flow paths 407, 408, 409, 410, 411, 412, and 413 connecting the chambers, and a port 109. The number of port 109 does not have to be one, and one port may be provided for each function. Furthermore, for example, the flow paths 413 and 412 may each have a dedicated port. The lysis chamber 101 holds a lysate 113 therein, and the washing solution chamber 105 holds a washing solution 111 therein.

[0101] As shown in Figure 11, a flow path 407 extending from the outlet of the lysis chamber 101 is connected to the inlet of the membrane chamber 108. A flow path 408 extending from the outlet of the membrane chamber 108 and a flow path 409 extending from the outlet of the washing solution chamber 105 join at a connection point (junction) 414 to form a flow path 410, which is connected to an inlet at the bottom (bottom surface: not necessarily the bottom surface) of the waste solution chamber 107. The flow path extending from the port 109 branches into flow paths 412 and 413 at a branch point 415. The flow path 413 is connected to the inlet of the washing solution chamber 105. The flow path 412 is connected to the top (top surface: not necessarily the top surface) of the waste solution chamber 107. Furthermore, the flow path 411 is also connected to the top (top surface: not necessarily the top surface) of the waste solution chamber 107.

[0102] As shown in FIG. 11, valve 401 is provided in flow path 407, valve 402 in flow path 409, valve 403 in flow path 410, valve 404 in flow path 411, valve 405 in flow path 412, and valve 406 in flow path 413.

[0103] <Purification Process> Fig. 12 is a diagram for explaining purification processes I to V according to Example 2. The flowchart of the purification process is the same as that shown in Fig. 7.

[0104] (i) Process I Process I shows the state during or immediately after the dissolution is completed in the sample dissolution procedure 202. At this time, valves 401, 402, 403, 404, 405, and 406 are closed.

[0105] (ii) Process II In process II, valves 402, 406, 403, and 404 are opened, and the washing solution 111 is pushed out of the washing solution chamber 105 at a pressure of 60 kPa or less for 30 seconds or less. At this time, air in flow path 409 moves to flow path 408 or 410, and flow path 409 is filled with the washing solution 111. Thus, discharging air from flow path 409 in process II is important for realizing continuous transport without trapping air between the lysate 113 and the washing solution 111.

[0106] (iii) Process III In Process III, valves 402 and 406 are closed, valves 401, 403, and 405 are opened, and 900 μL of lysate 113 is transported from lysis chamber 101 to waste chamber 107 via membrane chamber 108 at an applied pressure of −60 kPa for 2 minutes.

[0107] (iv) Process IV In process IV, once all of the lysate 113 has been transported to the membrane chamber 108, valves 403 and 405 are closed, valves 402 and 406 are opened, and 500 μL of wash solution 111 is transported from the wash solution chamber 105 to the lysis chamber 101 via the membrane chamber 108 at an applied pressure of 60 kPa for 3 minutes.

[0108] (v) Process V Process V shows a state in which the washing solution 111 is being transported from the washing solution chamber 105 to the lysis chamber 101 via the membrane chamber 108.

[0109] (vi) Process VI Process VI shows the state after the entire amount or a predetermined amount of the washing solution 111 has been transported from the washing solution chamber 105 to the lysis chamber 101 via the membrane chamber 108.

[0110] By adopting the above-described operation procedure, the lysate 113 and the washing solution 111 can be continuously transported without any air being trapped between the lysate 113 and the washing solution 111 .

[0111] It goes without saying that the pressure and time applied in Process III of Figure 12 are not limited to the values ​​described above. Furthermore, the combination of time and pressure must be such that 90% or more of the lysate 113 can be transported. Furthermore, the time and pressure required to transport almost all of the lysate 113 from the lysing chamber 101 to the purification membrane 102 may be confirmed by separate experiment and set in the implementation program.

[0112] A third embodiment will be described with reference to FIGS. 13, 14, and 15. FIG.

[0113] <Example of configuration of purification system 301 including main parts of flow path chip 114> Figure 13 is a diagram showing an example of configuration of a purification system 500 including main parts of the flow path chip 114 according to Example 3, which are the main parts of the flow path chip 114 when the sample dissolution procedure 202 is completed or is being performed.

[0114] The purification system 500 includes a lysis chamber 101, a washing solution chamber 105, a waste solution chamber 107, a membrane chamber 108, valves 501, 502, 503, 504, and 505, flow paths 506, 507, 508, 509, 510, and 511 connecting the chambers, and a port 109. The number of ports 109 does not have to be one; one port may be provided for each function. Alternatively, for example, dedicated ports may be provided for the flow paths 510 and 511, respectively. The lysis chamber 101 holds a lysate 113 therein, and the washing solution chamber 105 holds a washing solution 111 therein.

[0115] As shown in Figure 13, a flow path 506 extending from the outlet of the lysis chamber 101 is connected to one of two inlets (top or top surface; does not necessarily have to be the top surface) of the membrane chamber 108. A flow path 508 extending from the outlet of the membrane chamber 108 is connected to an inlet at the bottom (bottom surface; does not necessarily have to be the bottom surface) of the waste chamber 107. A flow path 507 extending from the outlet of the washing solution chamber 105 is connected to the other inlet of the membrane chamber 108. A flow path 511 extending from the port 109 is connected to the inlet of the washing solution chamber 105. Another flow path 510 extending from the port 109 is connected to the top (top surface; does not necessarily have to be the top surface) of the waste chamber 107. Furthermore, the flow path 509 is also connected to the top (top surface; does not necessarily have to be the top surface) of the waste chamber 107.

[0116] As shown in FIG. 13, valve 501 is provided in flow path 506, valve 502 in flow path 507, valve 503 in flow path 509, valve 504 in flow path 510, and valve 505 in flow path 511, respectively.

[0117] <Purification Process> Fig. 14 is a diagram for explaining purification processes I to VI according to Example 3. Fig. 15 is a flowchart corresponding to the purification process shown in Fig. 14.

[0118] (i) Process I Process I shows the state during or immediately after the completion of dissolution in the sample dissolution procedure 202 (step 1501). At this time, valves 501, 502, 503, 504, and 505 are closed.

[0119] (ii) Process II In process II, valves 501 and 504 are opened (step 1502), and 900 μL of lysate 113 is transported from lysis chamber 101 to waste chamber 107 via membrane chamber 108 at an applied pressure of −60 kPa for 2 minutes (step 1503).

[0120] (iii) Processes III and IV In Process III, once all of the lysate 113 has been transported to the membrane chamber 108, valve 504 is closed, valves 502 and 505 are opened (step 1504), and the cleaning solution 111 is transported from the cleaning solution chamber 105 to the flow path 506 via the membrane chamber 108 at an applied pressure of 30 kPa for 30 seconds (step 1505). At this time, air present in the space 108_1 on the inlet side of the membrane chamber 108 moves to the flow path 506, and the space 108_1 of the membrane chamber 108 is filled with the cleaning solution 111. Furthermore, the space 108_2 on the outlet side of the membrane chamber 108 is filled with the lysate 113. Thus, in Process IV, it is important to exhaust the air in the membrane chamber 108 to the flow path 506 in order to achieve continuous transport without trapping air between the lysate 113 and the cleaning solution 111.

[0121] (iv) Process V When the cleaning solution begins to enter the flow path 506, the valve 501 is closed and the valve 503 is opened (step 1506), and the cleaning solution 111 is transported from the cleaning solution chamber 105 to the waste chamber 107 via the membrane chamber 108 (step 1507).

[0122] (v) Process VI Process VI shows the state after the entire amount or a predetermined amount of the cleaning solution 111 has been transported from the cleaning solution chamber 105 to the waste chamber 107 via the membrane chamber 108.

[0123] By adopting the above-described operation procedure, the lysate 113 and the washing solution 111 can be continuously transported without any air being trapped between them.

[0124] A fourth embodiment will be described with reference to FIGS. 15, 16, and 17. FIG.

[0125] <Example of configuration of a purification system 600 including the main part of the flow path chip 114> Figure 16 is a diagram showing an example of configuration of a purification system 600 including the main part of the flow path chip 114 according to Example 4, which is the main part of the flow path chip 114 when the sample dissolution procedure 202 is completed or is being performed.

[0126] The purification system 600 includes a lysis chamber 101, a washing solution chamber 105, a waste chamber 107, a membrane chamber 108, valves 601, 602, 603, 604, 605, and 606, channels 607, 608, 609, 610, 611, 612, and 613 connecting the chambers, and a port 109. The number of ports 109 does not have to be one, and one port may be provided for each function. For example, the channels 612 and 613 may each have a dedicated port. The lysis chamber 101 holds a lysate 113 therein, and the washing solution chamber 105 holds a washing solution 111 therein.

[0127] As shown in Figure 16, a flow path 606 extending from the outlet of the lysis chamber 101 and a flow path 608 extending from the outlet of the washing solution chamber 105 join at a connection point (junction) 614 to form a flow path 609, which is connected to an inlet at the top (top surface: not necessarily the top surface) of the membrane chamber 108. A flow path 610 extending from an outlet provided at the bottom (bottom surface: not necessarily the bottom surface) of the membrane chamber 108 connects to an inlet at the bottom (bottom surface: not necessarily the bottom surface) of the waste chamber 107. A flow path 611 extending from an outlet provided at the top (top surface: not necessarily the top surface) of the membrane chamber 108 connects to the top (side surface: not necessarily the side surface, may be the top surface) of the waste chamber 107. A flow path 613 extending from the port 109 connects to the inlet of the washing solution chamber 105. Another flow path 612 extending from the port 109 is connected to the upper portion (top surface: not necessarily the top surface) of the waste liquid chamber 107. Furthermore, a flow path 615 is also connected to the upper portion (top surface: not necessarily the top surface) of the waste liquid chamber 107.

[0128] As shown in FIG. 16, valve 601 is provided in flow path 607, valve 602 in flow path 608, valve 603 in flow path 615, valve 604 in flow path 611, valve 605 in flow path 612, and valve 606 in flow path 613.

[0129] <Purification Process> Fig. 17 is a diagram for explaining purification processes I to V according to Example 4. The flowchart of the purification process is the same as that shown in Fig. 15 .

[0130] (i) Process I Process I shows a state during or immediately after the dissolution is completed in the sample dissolution procedure 202. At this time, valves 601, 602, 603, 604, 605, and 606 are closed.

[0131] (ii) Process II In process II, valves 601 and 605 are opened and 900 μL of lysate 113 is transported from lysis chamber 101 to waste chamber 107 via membrane chamber 108 at an applied pressure of −60 kPa for 2 minutes.

[0132] (iii) Process III In process III, after all of the lysate 113 has been transported to the membrane chamber 108, valves 601 and 605 are closed, and valves 602, 606, 604, and 603 are opened, and the wash solution 111 is transported from the wash solution chamber 105 through the membrane chamber 108 toward the flow path 611 at an applied pressure of 30 kPa for 30 seconds.

[0133] (iv) Process IV and Process V In Process IV, when the washing solution 111 enters the flow path 611, the valve 604 is closed and the washing solution 111 is transported from the washing solution chamber 105 to the waste chamber 107 via the membrane chamber 108. At this time, air present in the space 108_1 on the inlet side of the membrane chamber 108 moves to the waste chamber 107 via the flow path 611, and the space 108_1 of the membrane chamber 108 is filled with the washing solution 111. In addition, the space 108_2 on the outlet side of the membrane chamber 108 is filled with the lysate 113. Thus, in Process IV, it is important to discharge the air in the membrane chamber 108 to the waste chamber 107 via the flow path 611 in order to achieve continuous transport without trapping air between the lysate 113 and the washing solution 111.

[0134] (v) Process VI Process VI shows the state after the entire amount or a predetermined amount of the cleaning solution 111 has been transported from the cleaning solution chamber 105 to the waste chamber 107 via the membrane chamber 108.

[0135] By adopting the above-described operation procedure, the lysate 113 and the washing solution 111 can be continuously transported without any air being trapped between them.

[0136] Example 5 is a derivative of Example 1. The purification system 301' according to Example 5 has a configuration in which a valve 701 is added to the purification system 301 according to Example 1 (see FIG. 6).

[0137] <Refining Process> FIG. 18 is a diagram for explaining the refining processes I to V according to Example 5.

[0138] (i) Process I Process I shows the state during or immediately after the dissolution is completed in the sample dissolution procedure 202. At this time, valves 302, 303, 304, 305, and 701 are closed.

[0139] (ii) Process II In process II, valves 305, 304, and 701 are opened and 900 μL of lysate 113 is transported from lysis chamber 101 to waste chamber 107 via membrane chamber 108 at an applied pressure of −60 kPa for 1 minute.

[0140] (iii) Process III In process III, before air 702 following the transported lysate 113 passes the branch point (channel connection point (junction) 316) of channels 306 and 308 and channel 307, valves 304 and 701 are closed, valves 302 and 303 are opened, and washing solution 111 is transported from washing solution chamber 105 to lysis chamber 101 via channel 306 at an applied pressure of 30 kPa for 30 seconds. At this time, air in channel 307 moves to channel 308 or 306, and washing solution 111 is filled into channel 307. In process III, it is important to exhaust air from channel 307 in order to achieve continuous transport without trapping air between lysate 113 and washing solution 111.

[0141] (iv) Process IV In process IV, valves 315 and 701 are opened, and 500 μL of washing solution 111 is transported from washing solution chamber 105 to waste chamber 107 via membrane chamber 108 at 60 kPa for 3 minutes.

[0142] (v) Process V Process V shows the state after the entire amount or a predetermined amount of the cleaning solution 111 has been transported from the cleaning solution chamber 105 to the waste chamber 107 via the membrane chamber 108.

[0143] By adopting the above-described operation procedure, the lysate 113 and the washing solution 111 can be continuously transported without any air being trapped between them.

[0144] Example 6 is a derivative of Example 2. The purification system 400' according to Example 6 has a configuration in which a valve 801 is added to the purification system 400 according to Example 2 (see FIG. 11).

[0145] <Purification Process> FIG. 19 is a diagram for explaining purification processes I to V according to Example 6.

[0146] (i) Process I Process I shows the state during or immediately after the dissolution is completed in the sample dissolution procedure 202. At this time, valves 401, 402, 403, 404, 405, 406, and 801 are closed.

[0147] (ii) Process II In process II, valves 401, 403, 405, and 801 are opened and 900 μL of lysate 113 is transported from lysis chamber 101 to waste chamber 107 via membrane chamber 108 at an applied pressure of −60 kPa for 2 minutes.

[0148] (iii) Process III In Process III, once all of the lysate 113 has been transported to the membrane chamber 108, valves 405 and 801 are closed, valves 402, 406, and valve 404 are opened, and the washing solution 111 is pushed from the washing solution chamber 105 toward the waste chamber 107 at a pressure of 30 kPa or less for 30 seconds or less. At this time, air in flow path 409 moves to flow path 410 or the waste chamber 107. At this time, the air in flow path 409 moves to flow path 408 or 410, and the washing solution 111 is filled into flow path 409. Thus, in Process III, it is important to evacuate the air in flow path 409 in order to achieve continuous transport without trapping air between the lysate 113 and the washing solution 111.

[0149] (iv) Process IV In process IV, valve 403 is closed, valve 801 is opened, and 500 μL of washing solution 111 is transported from washing solution chamber 105 to lysis chamber 101 via membrane chamber 108 at 60 kPa for 3 minutes.

[0150] (v) Process V Process V shows the state after the entire amount or a predetermined amount of the washing solution 111 has been transported from the washing solution chamber 105 to the lysis chamber 101 via the membrane chamber 108.

[0151] By adopting the above-described operation procedure, the lysate 113 and the washing solution 111 can be continuously transported without any air being trapped between them.

[0152] (3) Supplementary explanations for each example: (i) In Example 1, in order to continuously transport the cleaning solution and the lysed product, it is necessary to switch from Process III to Process IV shown in Figure 6 at the correct timing. If the transport of the lysed product 113 is too long, air will enter the flow path 308, and if it is too short, a large amount of the lysed product 113 will remain in the lysing chamber 101, reducing the yield. For this reason, it is necessary to transport with high reproducibility or define the transport time using a liquid level detection sensor.

[0153] However, when the sample type is diverse or the viscosity of the liquid is high, the reproducibility of the transport of the lysate 113 is low. Furthermore, when a liquid level detection sensor is used, the number of components of the device increases, resulting in higher costs. There is also a risk that the liquid level detection sensor will not function properly.

[0154] On the other hand, in the switching from process III to process IV shown in Fig. 11 of Example 2, the switching from process II to process III shown in Fig. 14 of Example 3, and the switching from process II to process III shown in Fig. 17 of Example 4, the washing solution 111 and the lysate 113 can be continuously transported even if the time required to transport the lysate 113 exceeds the time required for transporting the lysate 113. Therefore, in these examples, a configuration can be realized that does not require a liquid level detection sensor while maintaining the yield, even if the liquid level detection sensor used in Example 1 is not used.

[0155] (ii) The lysate 113 contains macromolecules and particles derived from the sample, precipitates derived from the mixture with the lysis buffer, etc. These may get caught on the purification membrane during transport of the lysate 113, which may reduce the efficiency of transport of the wash solution 111 and eluate in the subsequent stages, or the efficiency of transport of fluids (air, nitrogen, and other gases) during drying.

[0156] In this regard, in the case of Example 3, the cleaning solution 111 is flowed from the opposite side of the lysate 113, which has the effect of pushing against any objects caught on the membrane during the transport of the lysate, thereby realizing smooth transport.

[0157] (iii) In the cases of Example 3, Example 4, Example 5 (a derivative of Example 1), and Example 6 (a derivative of Example 2), it is not necessary to divide the transfer of the cleaning solution 111 into multiple transfers. Therefore, it is not necessary to design the cleaning solution chamber 105 to accommodate multiple transfers. For example, a reagent transfer pack using an actuator, as shown in US 2006-134773 A1, may be used as the reagent chamber. However, in the cases of Example 5 (a derivative of Example 1) and Example 6 (a derivative of Example 2), if valve 701 or valve 801 is not provided, there is a possibility that fluid (air, nitrogen, or other gas) will move toward the membrane chamber 108. Therefore, although transfer is possible without valve 701 or valve 801, valve 701 or 801 can be provided to achieve more stable solution transfer. Furthermore, the flow path in Example 3 tends to be longer than in Example 1. Furthermore, the flow path in Example 4 requires an additional flow path compared to Example 1.

[0158] (4) Summary of Examples In each example, two types of solutions can be continuously delivered to the membrane-installed site in a channel with a purification membrane without installing a dedicated bubble removal device on the channel chip, thereby reducing the pressure required for solution delivery. This reduces or eliminates the number of gas-liquid interfaces with high Laplace pressure, and also reduces or eliminates the volume of fluid (air, nitrogen, or other gases) that must be delivered while the membrane is wet.

[0159] (i) According to Example 1 (see Figures 5 and 6), in a purification system (biomolecular analyzer) 301 in which a membrane chamber 108 is disposed between a lysis chamber (first chamber) 101, a washing chamber (second chamber) 105, and a waste chamber 107, a computer 115 controls the transport of a washing solution (second liquid) 111 at least until it passes a junction 316 between flow paths 306 to 309 (first flow paths) connecting the lysis chamber 101 to the waste chamber 107 and a flow path 307 (second flow path) extending from the washing chamber (second chamber) 105, and discharges fluid (air, nitrogen, or other gas) from the flow path 307 (second flow path). Next, the computer 115 controls the transport of a lysate (first liquid) 113 from the lysis chamber (first chamber) 101 to the waste chamber 107 via the membrane chamber 108. Then, a computer 115 controls the transport of the washing solution (second solution) 111 from the washing solution chamber (second chamber) 105 to the waste chamber 107 via the membrane chamber 108. This makes it possible to transport the lysate 113 and the washing solution 111 continuously to the waste chamber 107 without air being mixed between the lysate 113 and the washing solution 111 during solution transport. Since no fluid (air, nitrogen, or other gas) is mixed in, the pressure during solution transport can be reduced (see FIG. 10 ). This allows the use of flow paths with somewhat weak structures, thereby reducing the manufacturing cost of the purification system 301.

[0160] (ii) In Example 2 (see Figures 11 and 12), a membrane chamber 108 is disposed between a lysis chamber (first chamber) 101 and a washing solution chamber (second chamber) 105. The membrane chamber 108 and the washing solution chamber 105 are connected to a waste chamber 107. In the flow path of the purification system (biomolecular analyzer) 400, a computer 115 controls the transport of a washing solution (second liquid) 111 at least until the washing solution (second liquid) 111 passes through a junction 414 between a flow path 407-408 (first flow path) connecting the lysis chamber (first chamber) 101 to the waste chamber 107 and a flow path 409 (second flow path) extending from the washing solution chamber (second chamber) 105. The computer 115 then controls the transport of the lysate (first liquid) 113 from the lysis chamber (first chamber) 101 to the waste chamber 107 via the membrane chamber 108. The computer 115 controls the transport of the washing solution (second solution) 111 from the washing solution chamber (second chamber) 105 to the lysis chamber (first chamber) 101 via the membrane chamber 108. This prevents fluid (air, nitrogen, or other gas) from mixing between the lysate 113 and the washing solution 111 during solution transport, and after transporting the lysate 113 to the waste chamber 107, the washing solution 111 can be continuously transported to the lysis chamber 101. Since no fluid (air, nitrogen, or other gas) is mixed in, the pressure during solution transport can be reduced (see FIG. 10 as in Example 1). This allows the use of flow paths with somewhat weak structures, thereby reducing the manufacturing cost of the purification system 400.

[0161] (iii) According to Example 3 (see Figures 13 and 12), the membrane chamber has, on the upstream side, a first inlet connected to the lysis chamber (first chamber) 101, a second inlet connected to the washing solution chamber (second chamber) 105, and an outlet connected to the waste chamber 107; a flow path 506 (first flow path) extending from the lysis chamber (first chamber) 101 connects to the first inlet of the membrane chamber 108; a flow path 507 (second flow path) extending from the washing solution chamber (second chamber) 105 connects to the second inlet of the membrane chamber 108; and a flow path 508 (third flow path) extending from the outlet of the membrane chamber 108 connects to the waste chamber 107. In this flow path of the purification system (biomolecular analyzer) 500, the lysate (first liquid) 113 is transported from the lysis chamber (first chamber) 101 to the waste chamber 107 via the membrane chamber 108 by a computer 115. Next, the computer 115 transfers the washing solution (second solution) from the washing solution chamber (second chamber) 105 through the second inlet of the membrane chamber 108 to the space 108_1 between the first and second inlets of the membrane chamber 108 and the purification membrane, and in a state where the space 108_1 is filled with the washing solution (second solution) 111, the washing solution (second solution) 111 is discharged from the first inlet of the membrane chamber 108 to the flow path 506 (first flow path) side. Then, in a state where the washing solution (second solution) 111 is present on the flow path 506 (first flow path) side, the computer 115 transfers the washing solution (second solution) from the washing solution chamber (second chamber) through the membrane chamber 108 to the waste chamber 107. In this manner, it becomes possible to continuously transfer the lysate 113 and the washing solution 111 to the waste chamber 107 without mixing of fluids (air, nitrogen, other gases) between the lysate 113 and the washing solution 111 during solution transfer. Since no fluid (air, nitrogen, or other gas) is mixed in, the pressure during solution transport can be reduced (see FIG. 10 as in Example 1), so even if the flow path is somewhat weak in structure, it can be used, and the manufacturing cost of the purification system 500 can be reduced.

[0162] (iv) According to Example 4 (see Figures 16 and 17), a purification membrane 102, a membrane chamber 108 having an inlet connecting the lysis chamber (first chamber) 101 and the washing solution chamber (second chamber) 105 via a flow path 609, a communication port with the waste liquid chamber 107, and an outlet, flow paths 607 to 609 (first flow path) extending from the lysis chamber (first chamber) 101 and connecting to the inlet of the membrane chamber 108, and a flow path 614 extending from the washing solution chamber (second chamber) 105 to a junction 614 of the flow paths 607 to 609 (first flow path). In the flow paths of the purification system (biomolecular analyzer) 600, which includes a flow path 608 (second flow path) connected to the membrane chamber 101, a flow path 611 (communication flow path) connecting a communication port provided in the first space 108_1 between the inlet of the membrane chamber 108 and the purification membrane to the waste chamber 107, and a flow path 610 (third flow path) extending from the outlet of the membrane chamber 108 and connected to the waste chamber 107, a lysate (first liquid) 113 is transported from the lysis chamber (first chamber) 101 to the waste chamber 107 via the membrane chamber 108 by a computer 115. Next, a washing liquid (second liquid) 113 is transported from the washing liquid chamber (second chamber) 105 to the inlet of the membrane chamber 108 to the first space 108_1 and the flow path 611 (communication flow path). Then, with the first space 108_1 and the flow path 611 (communication flow path) filled with the washing solution (second solution) 111, the washing solution (second solution) 111 is transported from the washing solution chamber (second chamber) 105 to the waste chamber 107 via the membrane chamber 108 by the computer 115. In this manner, the lysate 113 and the washing solution 111 can be continuously transported to the waste chamber 107 without any fluid (air, nitrogen, or other gas) being mixed between the lysate 113 and the washing solution 111 during solution transport. Since no fluid (air, nitrogen, or other gas) is mixed, the pressure during solution transport can be reduced (see FIG. 10 as in Example 1). Therefore, even if the flow path is structurally weak, it can be used, and the manufacturing cost of the purification system 600 can be reduced.

[0163] 100 Biomolecule analysis device 101 Lysis chamber 102 Purification membrane 103 Reaction chamber 104 Lysis buffer chamber 105 Washing liquid chamber 106 Reagent chamber 107 Waste liquid chamber 108 Membrane chamber 109 Port 114 Flow path chip 115 Computer 301, 301', 400, 400', 500, 600 Purification system 313, 314 Liquid level detection sensor

Claims

1. A method for controlling liquid transport in a flow path of a biomolecule analyzer by a computer, comprising: The biomolecule analyzer includes a first chamber for accommodating a first liquid, a second chamber for accommodating a second liquid, a membrane chamber having a purification membrane, and a waste liquid chamber; The method comprises: controlling, by the computer, the second liquid to be transported at least past a confluence of a first flow path connecting the first chamber to the waste liquid chamber and a second flow path extending from the second chamber, and discharging a fluid different from the first and second liquids from the second flow path; controlling, by the computer, the transport of the first liquid from the first chamber to the waste liquid chamber via the membrane chamber; controlling, by the computer, the transport of the second liquid in the second chamber from the second chamber to the membrane chamber; A method comprising:

2. In claim 1, the membrane chamber is disposed between the first chamber, the second chamber and the waste chamber; The method further comprises: The method further comprises controlling, by the computer, the transport of the second liquid transported to the membrane chamber to the waste chamber.

3. In claim 1, the membrane chamber is disposed between the first chamber and the second chamber; The method further comprises: The method further comprises controlling, by the computer, the second liquid delivered to the membrane chamber to be further delivered to the first chamber.

4. In claim 1, The computer controls the applied pressure and the opening and closing of valves in response to input information on the flow rates of the first liquid and the second liquid, thereby transporting each of the first liquid and the second liquid.

5. In claim 4, the membrane chamber is disposed between the first chamber, the second chamber and the waste chamber; The computer (i) closes a first valve provided on the first flow path closer to the first chamber than the junction, and opens a second valve provided on the second flow path to apply a liquid delivery pressure, thereby transporting the second liquid at least past the junction; (ii) then closes the second valve, opens the first valve, and applies a liquid delivery pressure, thereby transporting the first liquid from the first chamber via the membrane chamber to the waste chamber; and (iii) further closes the first valve, reopens the second valve, and applies a liquid delivery pressure, thereby transporting the second liquid from the second chamber via the membrane chamber to the waste chamber.

6. In claim 4, the membrane chamber is disposed between the first chamber and the second chamber; The computer (i) closes a first valve provided between the first chamber and the membrane chamber, and opens a second valve provided in the second flow path and a third valve provided between the junction and the waste liquid chamber to apply a liquid delivery pressure, thereby transporting the second liquid at least until the second liquid passes the junction; (ii) then closes the second valve, opens the first and third valves to apply a liquid delivery pressure, thereby transporting the first liquid from the first chamber via the membrane chamber to the waste liquid chamber; and (iii) further closes the third valve, opens the first and second valves to apply a liquid delivery pressure, thereby transporting the second liquid from the second chamber via the membrane chamber to the first chamber.

7. In claim 4, the membrane chamber is disposed between the first chamber, the second chamber and the waste chamber; The computer (i) opens a first valve provided between the junction and the first chamber and a second valve provided between the junction and the membrane chamber, closes a third valve provided in the second flow path, and applies a liquid delivery pressure, thereby transporting the first liquid from the first chamber via the membrane chamber to the waste liquid chamber, thereby filling the area from the third valve to the inlet of the waste liquid chamber with the first liquid, (ii) then closes the second valve, opens the first and third valves, and applies a liquid delivery pressure, thereby transporting the second liquid at least until it passes the junction, and (iii) further closes the first and second valves, opens the third valve, and applies a liquid delivery pressure, thereby transporting the second liquid from the second chamber via the membrane chamber to the waste liquid chamber.

8. In claim 4, the membrane chamber is disposed between the first chamber and the second chamber; The computer (i) opens a first valve provided between the first chamber and the membrane chamber, a second valve provided between the membrane chamber and the junction, and a third valve provided between the junction and the waste liquid chamber, and closes a fourth valve provided in the second flow path to apply a liquid sending pressure, thereby transporting the first liquid from the first chamber to the waste liquid chamber via the membrane chamber, (ii) then transports the first liquid at least until it passes the junction, and (iii) then calculates a pressure between the first liquid outlet of the purification membrane and the first liquid outlet of the membrane chamber and the space between the first liquid outlet and the waste liquid chamber, and a flow path between the second valve and the second chamber is kept filled with the first liquid, the second valve is closed, and a liquid delivery pressure is applied by opening the third and fourth valves, thereby transporting the second liquid from the second chamber to the waste chamber and discharging a fluid contained in the second flow path and different from the first and second liquids into the waste chamber; and (iv) a liquid delivery pressure is applied by closing the third valve, and opening the first, second, and fourth valves, thereby transporting the second liquid from the second chamber to the first chamber via the membrane chamber.

9. In claim 1, The cross-sectional area of ​​the first flow path connected to the membrane chamber is 314 mm ２ The method is as follows.

10. In claim 1, The method, wherein the purification membrane is capable of retaining particles of 0.1 μm or larger.

11. In claim 1, The composition of the purification membrane is silica.

12. In claim 1, A method in which the first liquid contains at least one type of molecule selected from the group consisting of biopolymers including nucleic acids, proteins, lipids, or polysaccharides, or biomonomers including amino acids, lipids, sugars, or nucleic acid bases, and substances having molecules that contain derivatives of the biopolymers or the biomonomers in their structure.

13. In claim 1, The second liquid has a faster evaporation rate, a lower surface tension, or a larger contact angle with respect to the membrane than the first liquid.

14. In claim 1, The method, wherein the first liquid is a lysate, the second liquid is a wash liquid, and the fluid is air.

15. In claim 1, The method of claim 1, wherein the computer effects the delivery of the first and second liquids at a pressure less than a pressure required for the fluid to pass through the purification membrane after the first liquid has passed through the purification membrane.

16. In claim 4, The computer defines the timing of switching between the transport of the first liquid and the second liquid by a liquid level detection sensor, time, pressure, or a combination of these.

17. In claim 1, A method according to claim 1, wherein the volume of the second flow path is equal to or greater than the volume of a first space between the purification membrane and an inlet of the membrane chamber.

18. A method for controlling liquid transport in a flow path of a biomolecule analyzer by a computer, comprising: The biomolecule analyzer includes a first chamber for accommodating a first liquid, a second chamber for accommodating a second liquid, a membrane chamber having a purification membrane, and a waste liquid chamber, the membrane chamber having a first inlet, a second inlet, and an outlet, a first flow path extending from the first chamber is connected to the first inlet of the membrane chamber, a second flow path extending from the second chamber is connected to the second inlet of the membrane chamber, and a third flow path extending from the outlet of the membrane chamber is connected to the waste liquid chamber, The method comprises: transporting, by the computer, the first liquid from the first chamber through the membrane chamber to the waste chamber; transporting the second liquid from the second chamber through the second inlet of the membrane chamber to a first space between the first and second inlets of the membrane chamber and the purification membrane, and discharging the second liquid from the first inlet of the membrane chamber to the first flow path side while the first space is filled with the second liquid; transporting the second liquid from the second chamber to the waste liquid chamber via the membrane chamber while the second liquid is present on the first flow path side by the computer; A method comprising:

19. In claim 18, The computer controls the applied pressure and the opening and closing of valves in response to input information on the flow rates of the first liquid and the second liquid, thereby transporting each of the first liquid and the second liquid.

20. 20. In claim 19, The computer (i) opens a first valve provided in the first flow path and closes a second valve provided in the second flow path to apply a liquid delivery pressure, thereby transporting the first liquid from the first chamber to the waste chamber via the membrane chamber; (ii) then, while a second space between the purification membrane and the outlet of the membrane chamber is filled with the first liquid, opens the first and second valves to apply a liquid delivery pressure, thereby discharging the second liquid from the first inlet to the first flow path side while filling the first space of the membrane chamber with the second liquid; and (iii) further, closes the first valve and applies a liquid delivery pressure, thereby transporting the second liquid from the second chamber to the waste chamber via the membrane chamber, while the second liquid is present on the first flow path side.

21. A method for controlling liquid transport in a flow path of a biomolecule analyzer by a computer, comprising: The biomolecule analyzer includes a first chamber for accommodating a first liquid, a second chamber for accommodating a second liquid, a membrane chamber having a purification membrane, and a waste liquid chamber, a first flow path extending from the first chamber is connected to an inlet of the membrane chamber, a second flow path extending from the second chamber is joined to the first flow path at a joining point, and a communication flow path is further provided for connecting a first space between the inlet of the membrane chamber and the purification membrane to the waste liquid chamber, The method comprises: transporting, by the computer, the first liquid from the first chamber through the membrane chamber to the waste chamber; Transporting the second liquid from the second chamber through the inlet of the membrane chamber to the first space and the communication channel by the computer; transporting the second liquid from the second chamber to the waste liquid chamber via the membrane chamber while the first space and the communication channel are filled with the second liquid by the computer; A method comprising:

22. 22. In claim 21, The computer controls the applied pressure and the opening and closing of valves in response to input information on the flow rates of the first liquid and the second liquid, thereby transporting each of the first liquid and the second liquid.

23. In claim 22, The computer (i) opens a first valve provided in the first flow path and closes a second valve provided in the second flow path to apply a liquid delivery pressure, thereby transporting the first liquid from the first chamber via the membrane chamber to the waste liquid chamber; (ii) then, while a second space between the purification membrane and the outlet of the membrane chamber is filled with the first liquid, closes the first valve, opens a second valve provided in the second flow path and a third valve provided in the communication flow path to apply a liquid delivery pressure, thereby filling the first space of the membrane chamber and the communication flow path with the second liquid; and (iii) further closes the third valve and applies a liquid delivery pressure, thereby transporting the second liquid from the second chamber via the membrane chamber to the waste liquid chamber while maintaining a state in which the second liquid is present in the communication flow path.

24. A biomolecule analyzer having a flow path; A computer that controls liquid transport in the flow path, The biomolecule analysis device comprises: a first chamber containing a first liquid; a second chamber containing a second liquid; a waste chamber; a membrane chamber having a purification membrane and disposed between the first chamber, the second chamber and the waste chamber; Equipped with The computer includes: a process of controlling the transport of the second liquid at least past a confluence of a first flow path connecting the first chamber to the waste liquid chamber and a second flow path extending from the second chamber, and discharging a fluid different from the first and second liquids from the second flow path; a process of controlling the transport of the first liquid from the first chamber to the waste liquid chamber via the membrane chamber; a process of controlling the transport of the second liquid in the second chamber from the second chamber to the waste liquid chamber via the membrane chamber; Run the biomolecule purification system.

25. A biomolecule analyzer having a flow path; A computer that controls liquid transport in the flow path, The biomolecule analysis device comprises: a first chamber containing a first liquid; a second chamber containing a second liquid; a membrane chamber having a purification membrane and disposed between the first chamber and the second chamber; a waste chamber; The computer includes: a process of controlling the transport of the second liquid at least past a confluence of a first flow path connecting the first chamber to the waste liquid chamber and a second flow path extending from the second chamber, and discharging a fluid different from the first and second liquids from the second flow path; a process of controlling the transport of the first liquid from the first chamber to the waste liquid chamber via the membrane chamber; A process of transporting and controlling the second liquid in the second chamber from the second chamber to the first chamber via the membrane chamber; Run the biomolecule purification system.

26. A biomolecule analyzer having a flow path; A computer that controls liquid transport in the flow path, The biomolecule analysis device comprises: a first chamber containing a first liquid; a second chamber containing a second liquid; a membrane chamber having a purification membrane, a first inlet, a second inlet, and an outlet; a waste chamber; A first flow passage extending from the first chamber and connected to the first inlet of the membrane chamber; A second flow passage extending from the second chamber and connected to the second inlet of the membrane chamber; a third flow path extending from the outlet of the membrane chamber and connected to the waste liquid chamber; The computer includes: transporting the first liquid from the first chamber through the membrane chamber to the waste chamber; a process of transporting the second liquid from the second chamber through the second inlet of the membrane chamber to a first space between the first and second inlets of the membrane chamber and the purification membrane, and discharging the second liquid from the first inlet of the membrane chamber to the first flow path side while the first space is filled with the second liquid; a process of transporting the second liquid from the second chamber to the waste liquid chamber via the membrane chamber while the second liquid is present on the first flow path side; Run the biomolecule purification system.

27. A biomolecule analyzer having a flow path; A computer that controls liquid transport in the flow path, The biomolecule analysis device comprises: a first chamber containing a first liquid; a second chamber containing a second liquid; a waste chamber; a membrane chamber having a purification membrane, an inlet, a port communicating with the waste chamber, and an outlet; A first flow path extending from the first chamber and connected to the inlet of the membrane chamber; a second flow passage extending from the second chamber to a junction with the first flow passage; a communication channel that connects the communication port provided in a first space between the inlet of the membrane chamber and the purification membrane and the waste liquid chamber; a third flow path extending from the outlet of the membrane chamber and connected to the waste liquid chamber; A biomolecule purification system comprising:

28. 28. In claim 27, The computer includes: transporting the first liquid from the first chamber through the membrane chamber to the waste chamber; A process of transporting the second liquid from the second chamber through the inlet of the membrane chamber to the first space and the communication channel; a process of transporting the second liquid from the second chamber through the membrane chamber to the waste liquid chamber while the first space and the communication channel are filled with the second liquid; Run the biomolecule purification system.