Method for controlling liquid delivery in a flow path of a biomolecule analysis device with a computer and

Abstract

In order to prevent air from crossing a purification membrane without an additional flow path structure, the present invention proposes a method for controlling the delivery of a liquid in a flow path of a biomolecule analysis device with a computer, the biomolecule analysis device comprises a first chamber for accommodating a first liquid, a second chamber for accommodating a second liquid, a membrane chamber with a purification membrane, and a waste liquid chamber, and the method comprises the following steps: controlling by using a computer; a second liquid discharge section that conveys a second liquid to a position exceeding at least a confluence point of a first flow path connecting the first chamber and the waste liquid chamber and a second flow path extending from the second chamber, and discharges a fluid different from the first liquid and the second liquid from the second flow path; a computer is used for controlling, and the first liquid is conveyed from the first cavity to the waste liquid cavity through the membrane cavity; and delivering the second liquid of the second chamber from the second chamber to the membrane chamber under the control of the computer (fig. 6).

Description

Technical Field

[0001] The present disclosure relates to a method for controlling liquid delivery in a flow path of a biomolecule analysis device using a computer and a biomolecule purification system. Background Art

[0002] When performing gene analysis, for example, a process is used in which the sample is dissolved, the nucleic acid is purified / amplified as a preliminary process, and the amplified product is detected. This step involves the risk of contamination and cumbersome sample adjustment. Therefore, in the past, the sample was generally sent to an environment with complete experimental equipment such as a research institute, and the inspectors with specialized knowledge and technology adjusted and measured the sample and performed data analysis. However, problems such as the time spent on transporting the sample and the high equipment and personnel costs required to maintain the experimental equipment can be cited. In addition, when batch processing is used to improve efficiency, it is difficult to insert urgent samples.

[0003] In recent years, StoA (Sample-to-answer) systems that fully automatically implement everything from sample introduction to measurement and data acquisition have gradually emerged in various fields. In StoA systems, there are cases where flow path chips that integrate chambers, flow paths, and reagents are used. By using flow path chips, the following advantages can be obtained. That is, (i) non-experts can also easily measure, (ii) data can be obtained in a short time, (iii) devices with high mobility can be designed, (iv) errors caused by operation are reduced, and (v) reagents are easy to store.

[0004] As the application field of StoA flow path chip, potential applications include forensic medicine, in vitro diagnosis, identification of plant and animal species, biodefense, medicine, biotechnology, life sciences, defense, public health and agriculture. In the case of genetic analysis using StoA flow path chip, in order to prevent sample mixing between analyses, it is preferred that part or all of the flow path that directly contacts the sample is discarded after each measurement. In order to reduce the cost of disposable chips, countermeasures such as easy-to-manufacture design and the use of cheap materials can be cited.

[0005] However, in such a cheap chip, the pressure resistance of the chip is limited by the valve and the chip bonding strength. For example, in the case of Patent Document 1, the flow channel chip adopts a simple structure of bonding two pieces of thermoplastic resin. There is a record that the pressure resistance of the chip is limited by the valve and is 68kPa. In addition, for example, in the case of Patent Document 2, there is a record that the pressure resistance of the flow channel chip is, for example, 124kPa. In contrast, in the case of large-scale flow channel systems, such as liquid chromatographs, the pressure that can be used in solution delivery exceeds several MPa. Furthermore, in the case of spin columns widely used in the purification of nucleic acids, a maximum pressure of 500kPa can be applied. In this way, the pressure that can be used when implementing sample processing in the StoA flow channel chip tends to be lower than that of desktop equipment.

[0006] In the StoA system, since the delivery of the solution must be completed automatically with limited space and pressure, it is not desirable to have bubbles in the flow path. This is because there is a concern that the delivery of the solution may be incomplete or cause unexpected actions due to the mixing of air in the flow path. In order to solve such problems caused by the presence of air in the flow path, according to non-patent document 1, a material from which air can be discharged is used in the flow path. In addition, according to non-patent document 2 and patent document 3, a structure for discharging air is provided in the flow path. In addition, it can also be solved by applying high pressure, compressing or moving the air.

[0007] Prior art literature

[0008] Patent Literature

[0009] Patent Document 1: U.S. Patent No. 10233491

[0010] Patent Document 2: U.S. Patent No. 9354199

[0011] Patent Document 3: Japanese Patent No. 6613212

[0012] Non-patent literature

[0013] Non-patent document 1: "PDMS membranes with tunable gas permeability for microfluidic applications-RSC Advances (RSC Publishing) DOI: 10.1039 / C4RA1293B," RSC Adv., 2014, 4, 61415

[0014] Non-patent document 2: "Integrated Microfluidic System for Rapid Forensic DNAAnalysis: Sample Collection to DNAProfile / Analytical Chemistry (acs.org)," Anal.Chem.2010,82,16,6991-6999 Summary of the invention

[0015] Problems to be solved by the invention

[0016] In the purification and recovery of nucleic acids using purification membranes such as silicon oxide, a high recovery efficiency can be achieved with a simple flow path structure. However, when this purification method is implemented in a flow path with limited space for air to escape, the following problems arise. When air invades the chamber containing the membrane, the air must pass through the membrane chamber. For example, after the dissolved product passes through the membrane, when the air passes through the membrane while the membrane is wetted by the dissolved product, the pressure applied needs to exceed the Laplace pressure P defined by the following formula (1): L .

[0017] [Mathematical formula 1]

[0018]

[0019] 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 silica membrane with fine pores is used for purification, since d is small, it is believed that the Laplace pressure increases significantly.

[0020] In this regard, as described in the techniques disclosed in Patent Document 3, Non-Patent Document 1, and Non-Patent Document 2, by providing a structure for discharging air, it is possible to prevent air from passing through the membrane.

[0021] However, when this method is adopted, the flow path must have an additional structure, and there is a concern that there is a limitation on the material that can be used for the additional structure, or that the cost and size of the chip may increase due to the provision of the additional structure.

[0022] Furthermore, in the case of Patent Document 1, the purification membrane installed in the purification chamber does not cover the entire surface of the flow path. In this case, air passes over the side of the purification membrane, and the above-mentioned problem does not occur, but the proportion of dissolved products contacting the membrane decreases, so there is a concern that the recovery rate will decrease.

[0023] In view of such a situation, the present invention proposes a technology capable of preventing a fluid (air, nitrogen, or other gas) from passing over a purification membrane even without an additional flow path structure.

[0024] Technical solutions to solve problems

[0025] In order to solve the above-mentioned problems, the present disclosure, for example, proposes a method for controlling the liquid transport in the flow path of a biomolecule analysis device by a computer, characterized in that: the biomolecule analysis device 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, and the above-mentioned method includes the following processing: controlling by a computer to transport the second liquid to a position at least exceeding the confluence point of a first flow path connected from 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 liquid and the second liquid from the second flow path; controlling by a computer to transport the first liquid from the first chamber via the membrane chamber to the waste liquid chamber; and controlling by a computer to transport the second liquid of the second chamber from the second chamber to the membrane chamber.

[0026] Other features related to the present invention will be described in the description and drawings of this specification. In addition, the present invention is implemented by the combination of elements and multiple elements and the detailed description and the scope of rights of the additional claims. The description of this specification is only a typical example and does not limit the scope of rights or application examples of the present invention in any sense.

[0027] Effects of the Invention

[0028] According to the technology disclosed in the present invention, even without an additional flow path structure, it is possible to prevent air from passing over the purification membrane. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 1 is a diagram showing a configuration example of a biomolecule analysis device 100 according to the present embodiment.

[0030] Figure 2 It is a diagram showing a part of the biomolecule analyzer 100 and a derivative form of the flow channel chip 114 .

[0031] Figure 3 It is a diagram showing an example of a flow of bioanalysis using the biomolecule analyzer 100 .

[0032] Figure 4 This is a diagram showing an example of the internal structure of the computer 115.

[0033] Figure 5 1 is a diagram showing a configuration example of a purification system 301 including the main part of the flow channel chip 114 of Example 1, that is, the main part of the flow channel chip 114 when the sample lysing step 202 is completed or in progress.

[0034] Figure 6 It is a diagram showing purification processes I to V of Example 1.

[0035] Figure 7 is with Figure 6 The process shown corresponds to the flowchart.

[0036] Figure 8 It is a diagram for explaining the purification process of a comparative example.

[0037] Fig. 9 yes Figure 8 Flowchart of the purification process.

[0038] Fig.10 This is a diagram schematically showing the position of the solution during transportation in the experiment of Example 1 and the pressure change measured accordingly.

[0039] Fig.11 1 is a diagram showing a configuration example of a purification system 400 including the main part of the flow channel chip 114 of Example 2, that is, the main part of the flow channel chip 114 when the sample lysing step 202 is completed or in progress.

[0040] Fig.12 It is a diagram for explaining purification processes I to V of Example 2.

[0041] Fig.13 1 is a diagram showing a configuration example of a purification system 500 including the main part of the flow channel chip 114 of Example 3, that is, the main part of the flow channel chip 114 when the sample lysing step 202 is completed or in progress.

[0042] Fig.14 It is a diagram for explaining purification processes I to VI of Example 3.

[0043] Fig.15 is with Fig.14 Flowchart corresponding to the purification process shown.

[0044] Fig.16 1 is a diagram showing a configuration example of a purification system 600 including the main part of the flow channel chip 114 of Example 4, that is, the main part of the flow channel chip 114 when the sample lysing step 202 is completed or in progress.

[0045] Fig.17 It is a diagram for explaining purification processes I to V of Example 4.

[0046] Fig.18 It is a diagram for explaining purification processes I to V of Example 5.

[0047] Fig.19 It is a diagram for explaining purification processes I to V of Example 6. DETAILED DESCRIPTION

[0048] This embodiment proposes a technology that reduces the pressure required for liquid transport in the flow path chip and does not require structural robustness to set a purification membrane in the flow path chip through various examples. First, the various features of the biomolecule analysis device of the present embodiment are described and then the description of each embodiment is transferred to. In the drawings of the present application, there are cases where the same number represents the same functional elements. In addition, the drawings show specific embodiments and implementation examples that conform to the principles of the present disclosure, but these are used to understand the present disclosure and are not used to interpret the present disclosure in a limiting manner. In addition, in the present embodiment, the implementation of the present invention by those skilled in the art is fully described in detail, but it can also be changed to other implementations and methods, and it is necessary to understand that the structure, construction and replacement of various elements can be changed without departing from the scope and spirit of the technical idea of ​​the present disclosure. Therefore, the subsequent description should not be limited to this interpretation.

[0049] Furthermore, in the embodiments of the present disclosure, computer-controlled actions may be implemented using software running on a general-purpose computer, or may be implemented using dedicated hardware or a combination of software and hardware.

[0050] (1) Characteristics of Biomolecule Analysis Devices

[0051] <Flow channel chip>

[0052] In this embodiment, a "flow path chip (or simply referred to as a chip)" refers to a box that has reagents, chambers, and flow paths inside, and is disposable or can be used multiple times. The flow path chip may also have a power source for delivering the solution inside. In addition, a part or all of the reagents may be present in the chip. A part of the chamber may also have a temperature regulation function and a molecule capture function, a detection function, and a voltage application function.

[0053] The material of the flow channel chip is not particularly limited as long as it is a material generally used in the technical field. For example, as a material with a small amount of DNA adsorption, polypropylene and cycloolefin polymer (COP), cycloolefin copolymer (COC), polycarbonate, polyethylene terephthalate, polyurethane can be used. In addition, it is preferred to suppress the adsorption amount by applying a negatively charged modification on the surface. As other materials, for example, there can be mentioned metals such as gold, silver, copper, aluminum, tungsten, molybdenum, chromium, platinum, titanium, and nickel; alloys such as stainless steel, Hastelloy alloy, Inconel alloy, Monel alloy, and duralumin; silicon; glass materials such as glass, quartz glass, fused quartz, synthetic quartz, alumina, sapphire, ceramics, forsterite, and photosensitive glass; plastics such as polyester resin, polystyrene, polyethylene resin, ABS resin (Acrylonitrile Butadiene Styrene resin), polydimethylsiloxane (PDMS), nylon, acrylic resin, fluorine-containing resin, 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.

[0054] If the cross-sectional area of ​​the flow path is too large, there is a concern that liquid residue and sample loss may occur. In addition, it is difficult to apply pressure when the cross-sectional area is too large. On the other hand, if the cross-sectional area is too small, there are problems such as high pressure is required to transport the solution or the time spent on transportation becomes longer. Therefore, the cross-sectional area of ​​the flow path can be set to 1μm 2 ~314mm 2 As a more preferred structure, the cross-sectional area is set to 400 μm 2 ~100mm 2 As a further preferred structure, the cross-sectional area is set to 0.01 mm 2 ~10mm 2 range.

[0055] When the length of the flow path is too short, it is difficult to add elements such as valves, and when it is too long, there is a concern that the chip will become larger. Therefore, the length of the flow path connecting the structure of the chip can be set to 1μm to 100cm. As a more preferred range, it can be set to 1mm to 50cm. As a further preferred range, it can be set to 5mm to 30cm.

[0056] <Chamber>

[0057] The chamber is a space that can contain liquids or solids and has a diameter equal to or larger than that of the flow path. Reagents can be stored in the chamber, and PCR, dissolution, purification, etc. can also be performed in the chamber.

[0058] The capacity of the chamber is, for example, 0.01 μL to 10 L. When the capacity is larger than 10 L, it becomes difficult to transport the device. As a more preferred example, the capacity of the chamber can be set to 0.1 μL to 2 L. In addition, it is not necessary to set all the chambers in the chip, and they can also be provided in the device body or other independent chips.

[0059] When a cavity is provided in the chip, the capacity of the cavity can be set to 0.01 μL to 50 mL. If the capacity is larger than 50 mL, the chip becomes huge and difficult to store.

[0060] In the chip, one or more reagents are stored in one or more reagent storage chambers. When they are mixed at an undesirable time, performance degradation and other unexpected results may occur, so they are preferably separated by a valve, a film, air, or a flow path that is thin enough to prevent spontaneous mixing, or a separation mechanism composed of a combination thereof, just before use. In addition, long-term storage and chip mobility are achieved by isolating the reagents from the outside air.

[0061] Similarly, when the reagent is stored outside the chip (when a chamber is installed outside the flow channel chip), it is preferable to store the reagent in a state isolated from the outside air and separated from other purification system components by valves, membranes, air, etc.

[0062] <Purification system>

[0063] In this embodiment, the "purification system" refers to a system that uses a purification membrane in a flow channel chip and a membrane chamber that contains the purification membrane to capture biological molecules contained in a lysate contained in a lysate chamber, and then washes the lysate with a cleaning solution contained in a cleaning solution chamber.

[0064] The dissolution chamber and the cleaning liquid chamber may be provided outside the flow channel chip, or either of them may be provided inside the chip.

[0065] The purification system can be configured to selectively recover specific biomolecules. For example, it is possible to selectively extract DNA from a liquid containing proteins, DNA, ions, etc. In addition, it can also be a purification system for recovering molecules containing RNA, biopolymers (nucleic acids, proteins, lipids, polysaccharides), biomonomers (amino acids, lipids, sugars, nucleic acid bases) and their derivatives in the structure. In addition, it can also be a system for recovering multiple types of these molecules. As an example, there is a system that uses a silica membrane as a purification membrane and can recover DNA and RNA.

[0066] <Sample Type>

[0067] The sample provided by the purification system of this embodiment is not particularly limited as long as it is a sample of biological origin. The organism from which the sample is derived is not particularly limited, and samples derived from any organism such as vertebrates (e.g., mammals, birds, reptiles, fish, amphibians, etc.), invertebrates (e.g., insects, nematodes, crustaceans, etc.), plants, protozoa, plants, fungi, bacteria, viruses, etc. can be used.

[0068] Furthermore, when collecting a sample, a swab, filter paper, cloth, or the like can be used as a carrier, and each carrier can be introduced into a purification system.

[0069] <Dissolution products>

[0070] When the sample is transported to the membrane chamber, the sample needs to be in a form that can flow in the flow path. Therefore, when the state of the sample is solid (such as swab samples, etc.), it is preferred to dissolve or suspend the solid sample with a dissolution buffer to form a lysate composed of a fluid. The sample does not need to be completely dissolved, and the part that is still solid or has high viscosity after dissolution can be kept in the dissolution chamber. In addition, when the sample is a gas sample (such as air, exhaled breath, etc.), it is preferred to suspend the cells contained in the gas sample in a solvent to become a liquid sample. The modulation method for making the sample a lysate is conventionally carried out in the art, and those skilled in the art can easily understand it. For example, the dissolution buffer can contain chlorinated materials such as calcium hypochlorite. As another example, the substance can contain enzyme activities such as DNAase, RNAase, and protease. As needed, Chaotrope, surfactants, KOH, etc., substances that make biomolecules easy to be free, and substances that make nucleic acids easy to bind to purification membranes can also be added to the dissolution buffer. Heating, stirring, etc. can also be applied to the mixture as needed.

[0071] In this embodiment, the "lysate" refers to a substance obtained by making a biological sample into a liquid with a viscosity of 100,000 mPa·s or less using a lysis buffer. As a more preferred state, the lysate may have a viscosity of 10,000 mPa·s or less. As a further preferred state, the lysate may have a viscosity of 1000 mPa·s or less.

[0072] <Solution delivery control>

[0073] Valves can be used to control the delivery of solutions. Alternatively, solution control can be performed using flow path impedance.

[0074] In this embodiment (each example), air pressure is used as the power for transporting the solution. Alternatively, air pressure, mechanical compression, centrifugal force, etc. may be used as the power for transporting the solution.

[0075] When the pressure used for transportation is too high, it will exceed the pressure resistance of the chip. When it is too low, the time required for transportation will be longer, resulting in an extension of the measurement time. Therefore, the pressure used for transportation can be set to a range of 0.1kPa to 1MPa. The solution can be transported in a smaller range of 0.1kPa to 500kPa, and a further smaller range of 0.1kPa to 200kPa. In addition, regarding the time used for transportation, for each step, it can be implemented within a maximum of 1 hour, a smaller range of 10 minutes, and a further smaller range of 5 minutes. In addition, when the amount of liquid transported is too much, the time and pressure required for transportation increase, the cost of the chip increases, and the cost of the reagent also increases. Therefore, the amount of liquid transported can be set to within 1L for each reagent, a smaller range of 10mL, and a further smaller range of 2mL.

[0076] <Cleaning fluid>

[0077] In this embodiment, the "cleaning liquid" refers to a liquid used to wash away substances attached to the purification membrane that are not necessary in the subsequent process. In addition, the cleaning liquid may not be able to wash away all unnecessary substances, but may wash away some or all necessary substances.

[0078] According to the above formula (1), in the present embodiment, as a cleaning liquid, a cleaning liquid having the following characteristics can be used. First, it is preferred that the evaporation rate is faster than that of the dissolved product. In addition, it is preferred that the dissolved product and the cleaning liquid are miscible. Furthermore, it is preferred that the contact angle with the purification membrane is smaller (lower wettability) or the surface tension is lower than that of the dissolved product. As examples of cleaning liquids that meet such requirements, ethanol and isopropanol can be cited. In addition, a liquid containing 10% or more of these alcohols can be used as a cleaning liquid. In addition, a solution that does not meet the above conditions can also be used.

[0079] The number of cleaning liquids may be one or more. By using two or more cleaning liquids, cleaning can be performed more efficiently. In addition, they may be contained in one chamber or in two or more chambers. When two or more cleaning liquids are used, air may or may not be sandwiched between their conveyance. However, when the cleaning liquids after the second cleaning liquid have a faster evaporation rate or a smaller surface tension and a smaller contact angle with the purification membrane than the first cleaning liquid, it is preferred to convey them continuously.

[0080] <Purification membrane>

[0081] As the type of membrane, a silica membrane can be cited. As other examples of purification membranes, solid substrates with cellulose as the main component that can adsorb DNA, carboxylated particles, and ion exchange resins can be cited. In particular, a membrane having a hydroxyl group or a silica group on the surface can be used. The membrane can be any as long as it is a membrane that can hold particles of 100 μm or more. In addition, the preferred thickness is 1 μm or more. Furthermore, the finer the pores, the more efficiently the DNA can be recovered, so a membrane that can hold particles of 10 μm or more, more preferably 1 μm or more, and further preferably 0.1 μm or more can be used.

[0082] If the volume of the purification membrane is too small, the amount of biomolecules that can be adsorbed will decrease. On the other hand, if the volume is too large, there is a concern that the probability of unwanted molecular adsorption during purification or subsequent steps and the transport efficiency of the solution will deteriorate. The area used in the examples described below is 12.5 mm 2 However, for example, 1 mm 2 ~314mm 2 The size of the membrane is not limited.

[0083] <Detection method>

[0084] PCR is used for amplification in the downstream of this purification system. After amplification, CE (Capillary Electrophoresis) is used for detection. As other examples, MPS (Massively parallel sequencing), pyrophosphate sequencing, Sanger sequencing, nanopore sequencing, chromatography, electrical measurement, spectroscopy, NMR, RFLP (Restriction Fragment Length Polymorphisms) etc. can also be used.

[0085] <Other Supplementary Matters>

[0086] In all the figures used to illustrate the present embodiment, the same figure mark is added to the parts with the same function, and its repeated description is omitted as much as possible. Below, each embodiment of the present embodiment is described in detail based on the drawings. The measurement method, the structure of the device, the type of substance and the material recorded in each embodiment are an example for making the idea of ​​the present embodiment concrete, and do not strictly specify the material and size of the measurement principle and the device. In addition, the specific pressure value recorded in each embodiment is an example for making the idea of ​​the present embodiment concrete, and is not strictly specified. Furthermore, the specific sample type recorded in each embodiment, and the composition and liquid volume of the purification kit are an example for making the idea of ​​the present embodiment concrete, and are not an example for strictly defining the chemical composition and time. In addition, the specific type of the measured object recorded in each embodiment and the type of solution and its concentration are an example for making the idea of ​​the present embodiment concrete, and are not an example for strictly defining the chemical composition.

[0087] <Configuration example of biomolecule analysis device>

[0088] Figure 1 1 is a diagram showing a configuration example of a biomolecule analyzer 100 according to the present embodiment. The biomolecule analyzer 100 includes a flow channel chip 114 and a computer 115 for performing biomolecule analysis.

[0089] The flow path chip 114 has a dissolution chamber 101 for introducing and dissolving the sample, a membrane chamber 108 for storing a purification membrane 102, a reaction chamber 103 for performing DNA amplification, a waste liquid chamber 107, and a port 109 connected to the outside of the chip for fluid flow. In addition, the configuration of each chamber in the flow path chip 114 and the flow path connection between the chambers are different in each embodiment, so the chamber configuration and flow path connection and the transportation operation of the dissolved product and the cleaning liquid will be described later.

[0090] As the purification membrane 102, a glass fiber membrane GF / F sold by Whatman Co., Ltd. cut into a membrane with a diameter of 4 mm is provided. Other membranes may also be used as the purification membrane 102. In addition, the diameter of the purification membrane (the length in the long side direction in the case of a rectangle) can be set to a range of 0.1 mm to 100 mm.

[0091] The solution is transported by applying pressure through port 109, and reagents and amplification products can be exchanged with the outside of the chip. In addition, the pressure can be applied using a pressure generating device provided outside the chip. Figure 1The flow path chip 114 adopts a structure that includes chambers 104, 105 and 106 for storing reagents and the like. In addition, regarding the membrane chamber 108, the capacity of the portion close to the dissolution chamber 101 in the space separated by the membrane is 10μL, and the capacity of the portion close to the waste liquid chamber 107 is 10μL. The dissolution buffer chamber 104 stores a dissolution buffer 110 for dissolving the sample, the cleaning liquid chamber 105 stores a cleaning liquid 111, and the reagent chamber 106 stores an eluate or a reagent 112 used in the reaction. In addition, part of the above functions may also be integrated in the same chamber. The positional relationship of the above chip components is not limited to Figure 1 , the flow path can also be connected with Figure 1 different.

[0092] Figure 2 It is a diagram showing a part of the biomolecule analyzer 100 and a derivative form of the flow channel chip 114 . Figure 2 In the embodiment, the dissolution buffer chamber 104, the cleaning liquid chamber 105 and the waste liquid chamber 107 are arranged outside the flow channel chip 114. By adopting such a method, the advantages of miniaturization and cost reduction of the flow channel chip 114 can be obtained.

[0093] <Example of Bioanalysis Process>

[0094] Figure 3 1 is a diagram showing an example of a process of performing biological analysis using the biological molecule analyzer 100. Figure 3 As shown, the biological analysis includes, for example, a sample receiving step 201 , a sample dissolving step 202 , a sample generating step 203 , a sample amplifying step 204 and a sample detecting step 205 .

[0095] In the sample receiving step 201 , before or after the sample is placed in the lysis chamber 101 , the lysis buffer 110 is supplied from the chamber 104 to the lysis chamber 101 (a step of introducing the sample into the lysis chamber 101 ).

[0096] Next, the sample dissolution step 202 starts dissolution. In the sample generation process 203, the lysate 113 is transported from the dissolution chamber 101 to the purification membrane 102, and the DNA is bound to the membrane and purified. After purification, the step of drying the washing solution and the like can be entered. The dissolved DNA is transported to the reaction chamber 103.

[0097] In the sample amplification step 204, the purified DNA is amplified. In the sample detection step 205, the amplified DNA is measured. The sample detection step 205 can be performed in the flow channel chip 114, or can be transported to a detection unit outside the flow channel chip 114 and performed in a separately provided measurement unit.

[0098] In addition, steps 201 to 205 may be performed in parallel. In addition, some steps may be skipped, or other steps may be added.

[0099] <Internal Configuration Example and Control Operation of Computer 115>

[0100] Figure 4 115 is a diagram showing an example of the internal structure of the computer 115. The computer 115 includes a processor (not shown), a user interface 1151, and a database (storage device) 1152.

[0101] The user interface 1151 includes an input screen and an output screen, and can receive and communicate with the user. Figure 3 Parameters related to the implementation process of the process, such as the time and temperature of each step, pressure, flow rate, process, etc., are stored in the database 1152. In addition, various parameters can be stored in advance in the database 1152.

[0102] The computer 115 can open and close the valve of the flow channel chip 114, control the temperature, and control the pressure / flow rate based on the various parameters recorded in the database 1152. In addition, the computer 115 can automatically control Figure 3 In addition, the user can also assist and implement Figure 3 Part of the implementation steps shown.

[0103] (2) Embodiment

[0104] Example 1

[0105] refer to Figures 5 to 7 , Example 1 is described below.

[0106] <Configuration Example of the Purification System 301 Having Main Parts of the Flow Channel Chip 114>

[0107] Figure 5 1 is a diagram showing a configuration example of a purification system 301 including the main part of the flow channel chip 114 of Example 1, that is, the main part of the flow channel chip 114 when the sample lysing step 202 is completed or in progress.

[0108] The purification system 301 includes a dissolution chamber 101, a cleaning liquid chamber 105, a waste liquid 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 port 109 may not be one, but may be provided for each function. For example, dedicated ports may be added to the flow paths 311 and 310, respectively.

[0109] The dissolution chamber 101 holds the dissolved product 113 inside, and the cleaning liquid chamber 105 holds the cleaning liquid 111 inside. In addition, the liquid level detection sensor 313 is provided in the waste liquid chamber 107, and the liquid level detection sensor 314 is provided in the dissolution chamber 101. More than one liquid level detection sensor may be provided, or no liquid level detection sensor may be provided. In addition, liquid level detection sensors may also be provided in other chambers such as the dissolution chamber 101 and the waste liquid chamber 107.

[0110] like Figure 5 As shown, the flow path 306 extending from the outlet of the dissolution chamber 101 and the flow path 307 extending from the outlet of the cleaning liquid chamber 105 converge at a connection point (confluence point) 316 to form a flow path 308 and connect to the inlet of the membrane chamber 108. The flow path 309 extending from the outlet of the membrane chamber 108 is connected to the inlet of the lower part (bottom surface: it may not be the bottom surface) of the waste liquid chamber 107. In addition, two flow paths 310 and 311 are extended from the port 109, and the flow path 310 is connected to the upper part (top surface: it may not be the top surface) of the waste liquid chamber 107, and the flow path 311 is connected to the inlet of the cleaning liquid chamber 105. Furthermore, the flow path 312 is connected to the upper part (top surface: it may not be the top surface) of the waste liquid chamber 107.

[0111] In addition, if Figure 5 As shown, valve 302 is provided in flow path 307, valve 303 is provided in flow path 311, valve 304 is provided in flow path 310, valve 305 is provided in flow path 306, and valve 315 is provided in flow path 312.

[0112] <Purification process>

[0113] Figure 6 It is a diagram showing purification processes I to V of Example 1. In addition, Figure 7 is with Figure 6 The process shown corresponds to the flowchart. In addition, the control of the delivery of the dissolved product 113 and the cleaning solution 111 by opening and closing each valve and applying pressure in each process is performed by the computer (processor) 115.

[0114] (i) Process I

[0115] Process I indicates a state during or immediately after dissolution in the sample dissolution step 202. At this time, valves 302, 303, 304, 305, and 315 are closed (step 701).

[0116] (ii) Process II

[0117] In process II, valves 302, 303, and 315 are opened (step 702), and the cleaning liquid 111 is squeezed out from the cleaning liquid chamber 105 to the membrane chamber 108 at a pressure of 30 kPa or less in 30 seconds or less (step 703). At this time, the air in the flow path 307 moves to the flow path 308 or 306, and the cleaning liquid 111 is filled in the flow path 307. Exhausting the air in the flow path 307 in process II is important for achieving continuous transportation without air being trapped between the dissolved product 113 and the cleaning liquid 111.

[0118] (iii) Process III

[0119] In process III, valves 302, 303, and 315 are closed, and valves 305 and 304 are opened (step 704), and 900 μL of the dissolved product 113 is delivered from the dissolution chamber 101 to the waste liquid chamber 107 via the membrane chamber 108 in 1 minute by applying a pressure of -60 kPa (step 705). The delivery amount (900 μL) of the dissolved product 113 is controlled by the computer 115 based on the liquid level detection sensor 314 and the application time (1 minute) of the specified pressure (60 kPa).

[0120] (iv) Process IV

[0121] In process IV, before the air following the dissolved product 113 exceeds the branch point (connection point (confluence point) 316) of the flow paths 306 and 308 and the flow path 307, the valves 304 and 305 are closed, and the valves 302, 303, and 315 are opened (step 706), and 500 μL of the cleaning liquid 111 is delivered from the cleaning liquid chamber 105 to the waste liquid chamber 107 via the membrane chamber 108 in 3 minutes by applying a pressure of 60 kPa (step 707). The delivery amount (500 μL) of the cleaning liquid 111 is controlled by the computer 115 based on the liquid level detection sensor 314 and the application time (3 minutes) of the specified pressure (60 kPa).

[0122] By executing the operations shown in processes I to IV, it is possible to realize continuous transfer without trapping air between the dissolved product 113 and the cleaning liquid 111 .

[0123] <Comparative Example>

[0124] Figure 8 It is a diagram for explaining the purification process of a comparative example. Fig. 9 yes Figure 8 Flowchart of the purification process.

[0125] (i) Process I

[0126] Process I represents a state during or immediately after the dissolution in the sample dissolution step 202 (step 901). At this time, valves 302, 303, 304, and 305 are closed.

[0127] (ii) Process II

[0128] In process II, valves 304 and 305 are opened (step 902 ), and the lysate 113 is transported from the lysis chamber 101 to the waste chamber 107 via the membrane chamber 108 (step 903 ).

[0129] (iii) Process III

[0130] In process III, after the delivery of the dissolved product 113 is completed, the valves 305 and 304 are closed (step 904).

[0131] (iv) Process IV

[0132] In process IV, valves 302, 303, and 315 are opened (step 904). Cleaning liquid 111 is delivered from the cleaning liquid chamber 105 to the waste liquid chamber 107 via the membrane chamber 108 (step 905).

[0133] (v) Process V

[0134] Process V represents a state after the entire amount or a predetermined amount of the cleaning solution 111 is transferred to the waste liquid chamber 107 via the membrane chamber 108 .

[0135] In the purification process of this comparative example, air present in the flow paths 307 and 308 enters between the solution product 113 and the cleaning solution 111. Therefore, the cleaning solution 111 needs to exceed the Laplace pressure generated between the solution product 113 and the purification membrane 102 in order to reach the membrane.

[0136] <About technical effects>

[0137] In order to confirm the technical effect of the dissolution product and cleaning solution delivery action of Example 1, a comparative experiment was conducted using a simple flow path system for the pressure required for solution delivery when air enters the flow path and when air does not enter the flow path. In addition, the technical effect is discussed here based on the experimental results of Example 1, but the technical effects cited here are also the same for Examples 2 to 4 described later and derivative forms of the embodiments.

[0138] (i) Implementation conditions

[0139] In the flow system, the lysate and QIK cleaning solution (AW1) adjusted according to the protocol of Qiagen's QIK (Qiaamp investigator kit DNA) were used. The lysate and cleaning solution were placed in 1.5 mL tubes, pressurized with a syringe pump, and transported to the polycarbonate membrane chamber. The volume of the solution that the membrane chamber can hold is about 20 μL, and the center is divided by a membrane, and the capacity of the upstream and downstream is about 10 μL. The liquid contact area of ​​the purification membrane is set to 3.1 mm 2 The waste liquid tube set at the rear of the membrane chamber was used to recover the dissolved product and the washing liquid after passing through the membrane chamber. The extrusion speed of the syringe pump was set to 4 mL / min.

[0140] In the transport (process A) of the process (comparative example) other than Example 1, 100 μL of dissolved product, 100 μL of air, and 500 μL of cleaning solution (QIK cleaning solution AW1) were sequentially transported to the membrane chamber, and the time change of pressure was measured. In order to reproduce the transport method of Example 1, in process B, 100 μL of dissolved product and 500 μL of cleaning solution were continuously transported without air. The membrane used was a glass fiber membrane GF / F from Whatman.

[0141] (ii) Experimental results

[0142] Fig.10 This is a diagram schematically showing the position of the solution during transportation in the above-mentioned experiment and a diagram showing the pressure change measured accordingly. Fig.10 In the figure, the dotted line graph represents the result of process A (comparative example), and the solid line graph represents the result of process B (example 1).

[0143] The extrusion speed of the syringe pump is faster than the movement speed of the solution, so the pressure rises with time while the dissolved product passes through the membrane, reaching 20 kPa ( Fig.10 (solid line, dashed line) steps (a)→(b)).

[0144] In the case of process A, when the dissolved product completely permeates and air enters the membrane, the liquid level does not change. Furthermore, when the syringe pump is continuously pushed, air slowly begins to pass through the membrane at a pressure of more than 80 kPa. When the pressure reaches 180 kPa, the cleaning liquid reaches the membrane chamber ( Fig.10 (dashed line) Step (c) → (d)). After the cleaning liquid passed through the membrane and completely permeated, it was confirmed that the pressure dropped sharply to around 30 kPa ( Fig.10 (dashed line) step (d)).

[0145] In the case of process B, when the dissolved product completely permeates and the cleaning solution enters the membrane, the solution does not stop ( Fig.10(Solid line) Step (b) → (c)). The pressure did not exceed 40 kPa until the cleaning liquid completely penetrated. In addition, the necessary pressure for transporting the cleaning liquid alone was confirmed, and the transport was completed within 40 kPa (no data recorded).

[0146] (iii) Experimental results

[0147] As described above, based on the experimental results of Flow A (Comparative Example) and Flow B (Example 1), it was confirmed that the required pressure can be reduced to 1 / 3 by using the conveying method of Example 1 (the same applies to other Examples described below).

[0148] The reason why high pressure is not required when air enters after the cleaning liquid has passed through is that the evaporation rate of the cleaning liquid is much higher than that of the dissolved product. More than 50% of the volume of the cleaning liquid (QIK, AW1) is composed of ethanol.

[0149] In this embodiment, the maximum pressure required for conveying the two liquids is reduced by continuously conveying the lysate and the cleaning liquid. The effect of this embodiment is not limited to the above-mentioned reagent kit, and the composition of the lysate or the cleaning liquid may also be different. As a combination of the first liquid (lysate) and the second liquid that is more likely to obtain the effect of pressure reduction when the second liquid (cleaning liquid) is completely permeated before the sample detection step 205, the following elements can be cited.

[0150] (a) the second liquid evaporates faster than the first liquid;

[0151] (b) the second liquid has a lower surface tension than the first liquid;

[0152] (c) the second liquid has a larger contact angle with the membrane than the first liquid;

[0153] (d) The second liquid has a lower viscosity than the first liquid.

[0154] When the main component of the first liquid is water, as the second liquid satisfying one or more of the above conditions, a solution containing 10% or more of an alcohol having 4 or less carbon atoms can be cited. More preferably, a solution containing 30% or more of an alcohol having 3 or less carbon atoms can be cited. Even more preferably, a solution containing 40% or more of ethanol can be cited. In addition, the second liquid can be appropriately selected according to the composition of the first liquid, and the first liquid can be appropriately selected according to the composition of the second liquid.

[0155] According to Example 1 (experimental results) of the present disclosure, the necessary pressure can be reduced. Then, by reducing the necessary pressure, a high-density purification membrane can be set in a structurally fragile flow path chip. Here, as a structurally fragile chip, there can be cited a chip using a valve and a chip bonded together or a soft material. The manufacture of a structurally fragile chip has the advantages of reducing chip costs and improving chip performance. In addition, by reducing the necessary pressure, chemical reactions that are hindered by applying high pressure can be carried out efficiently. Furthermore, by applying high pressure, the measurement of biological molecules can be facilitated.

[0156] In addition to reducing pressure, the technology disclosed herein can shorten the time spent on transporting solutions. By using the technology disclosed herein, the membrane is prevented from drying after the solute passes through and before cleaning, thereby enjoying advantages such as increased biomolecule yield and increased cleaning efficiency.

[0157] (iv) Membranes for DNA recovery

[0158] In the biomolecule analysis device 100, by providing a high-density membrane, efficient DNA recovery can be achieved. As an indicator of the fineness of the pores, there is a "particle retention size". This means that particles with a particle size greater than the value recorded in the particle retention size are retained by the membrane. That is, the smaller the value, the finer the pores of the membrane. The membrane used as a purification membrane can use a membrane with a particle retention size of 100 μm or more. For example, it is preferred to use a membrane with finer pores than the "Fusion5" membrane sold by Whatman. In addition, as a membrane capable of more efficiently recovering DNA, the "GF / D" membrane sold by Whatman can be cited. The particle retention size of this membrane is 2.7 μm. In addition, as a membrane capable of more efficiently recovering DNA, the "GF / F" membrane sold by Whatman can be cited. The particle retention size of this membrane is 0.7 μm.

[0159] In addition, by overlapping multiple membranes, efficient DNA recovery can be achieved. For example, in the case of GF / F, by overlapping more than 2 membranes, DNA can be recovered more efficiently than with only one purification. The thickness of the membrane is preferably more than 0.001 mm. In the case of GF / F, the thickness of the membrane is 0.6 mm.

[0160] (v) Regarding the membrane chamber 108

[0161] The volume of the membrane chamber 108 is less than 30% of the volume of the dissolution chamber 101 or the cleaning liquid chamber 105. The cross-sectional area of ​​the flow path through which the two liquids pass in common and in the direction perpendicular to the advancing direction is less than 50% of the maximum cross-sectional area of ​​the chamber, and the volume of the flow path through which the two liquids pass in common is less than 50% for each of the two liquids. Therefore, the lysate and the cleaning liquid will not mix more than 50% before the waste liquid chamber. Therefore, even if the solution is transported using the technology disclosed in the present invention, the binding efficiency of DNA will not be significantly reduced.

[0162] The size of the membrane chamber 108 is 0.1 μL to 1 mL. The smaller the membrane chamber 108 is, the smaller the loss during elution is, so the size of the membrane chamber 108 is more preferably 0.1 μL to 100 μL, and further preferably 0.1 μL to 30 μL.

[0163] (vi) About flow path

[0164] The volume of the flow path 307 is 0.1 μL to 1 L (the volume of the flow path increases when a longer flow path is used). When the volume of the flow path 307 is larger than 1 L, it takes time to transport the solution. On the contrary, when the volume is smaller, it is necessary to narrow the flow path width or reduce the distance between the chambers, and the degree of freedom of design is reduced. In addition, when there is a chamber outside the chip, a larger chamber is preferred. When the flow path is too thin, there is a concern that unexpected actions such as blockage of the flow path may occur. The more preferred volume of the flow path is 1 μL to 100 mL, and in other embodiments it is 30 μL to 100 mL.

[0165] The volume of the flow path 307 can be set to be equal to or larger than the volume of the space 108_1 above the purification membrane 102 in the membrane chamber 108 .

[0166] In addition, when drying is performed after purification, it is preferable to ensure that the flow path is thick to a certain extent. In this case, the thickness of the flow paths 302 and 303 is preferably 0.03 mm. 2 .

[0167] <About process switching control>

[0168] (i) Control using a liquid level detection sensor

[0169] Figure 6 The liquid level detection sensor can be used in the switching of processes I, II, III, IV and V. For example, when switching from process III to IV, liquid level detection sensors 313 and 314 are used. As liquid level detection sensors 313 and 314, sensors that detect electrically, optically, acoustically, or mechanically can be used.

[0170] When the liquid level detection sensor 313 is used, when switching from process III to IV, the signal of the liquid level detection sensor 313 that detects that the liquid level of the waste liquid chamber 107 exceeds that of the liquid level detection sensor 313 can be used. However, the liquid level detection sensor 313 needs to be set to be able to detect the position of the liquid level of the waste liquid chamber 107 when the remaining amount of the dissolved product 113 in the dissolution chamber 101 is less than 10%.

[0171] In addition, when the liquid level detection sensor 314 is used, when switching from process step II to III, a signal detected when the liquid level in the dissolution chamber is lower than that of the liquid level detection sensor 314 may be used. However, the liquid level detection sensor 314 needs to be set to be able to detect the position of the liquid level when the remaining amount of the dissolved product 113 in the dissolution chamber 101 is less than 10%.

[0172] (ii) Control based on pressure, time, etc.

[0173] Figure 6 In the switching of processes I, II, III and IV, a pre-set pressure and time may also be used. In addition, the pressure and time may also be adjustable by the operator. Furthermore, a mechanism for measuring viscosity may also be provided in the dissolution chamber 101 or the like.

[0174] In addition, flow sensors and pressure sensors can be placed in specific locations to define the timing of process switching.

[0175] Example 2

[0176] refer to Figure 7 , Fig.11 and Fig.12 , Example 2 is described below.

[0177] <Configuration Example of the Purification System 301 Having Main Parts of the Flow Channel Chip 114>

[0178] Fig.11 1 is a diagram showing a configuration example of a purification system 400 including the main part of the flow channel chip 114 of Example 2, that is, the main part of the flow channel chip 114 when the sample lysing step 202 is completed or in progress.

[0179] The purification system 400 includes a dissolution chamber 101, a cleaning liquid chamber 105, a waste liquid 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 port 109 may not be one, and one port may be provided for each function. In addition, for example, dedicated ports may be added to the flow paths 413 and 412, respectively. The dissolution chamber 101 holds a dissolved product 113 inside, and the cleaning liquid chamber 105 holds a cleaning liquid 111 inside.

[0180] like Fig.11 As shown, the flow path 407 extending from the discharge port of the dissolution chamber 101 is connected to the inlet of the membrane chamber 108. The flow path 408 extending from the discharge port of the membrane chamber 108 and the flow path 409 extending from the discharge port of the cleaning liquid chamber 105 converge at a connection point (confluence point) 414 to form a flow path 410 and is connected to the inlet of the lower part (bottom surface: it may not be the bottom surface) of the waste liquid chamber 107. The flow path extending from the port 109 is divided into a flow path 412 and a flow path 413 at a branch point 415. The flow path 413 is connected to the inlet of the cleaning liquid chamber 105. The flow path 412 is connected to the upper part (top surface: it may not be the top surface) of the waste liquid chamber 107. Furthermore, the flow path 411 is also connected to the upper part (top surface: it may not be the top surface) of the waste liquid chamber 107.

[0181] In addition, if Fig.11 As shown, valve 401 is provided in flow path 407, valve 402 is provided in flow path 409, valve 403 is provided in flow path 410, valve 404 is provided in flow path 411, valve 405 is provided in flow path 412, and valve 406 is provided in flow path 413.

[0182] <Purification process>

[0183] Fig.12 Schematic diagram for explaining purification processes I to V of Example 2. Figure 7 same.

[0184] (i) Process I

[0185] Process I indicates a state during or immediately after dissolution in the sample dissolution step 202. At this time, valves 401, 402, 403, 404, 405, and 406 are closed.

[0186] (ii) Process II

[0187] In process II, valves 402, 406, 403, and 404 are opened, and the cleaning liquid 111 is squeezed out from the cleaning liquid chamber 105 at a pressure of 60 kPa or less in 30 seconds or less. At this time, the air in the flow path 409 moves to the flow path 408 or 410, and the cleaning liquid 111 is filled in the flow path 409. In this way, exhausting the air in the flow path 409 in process II is important for achieving continuous conveyance without air being trapped between the dissolved product 113 and the cleaning liquid 111.

[0188] (iii) Process III

[0189] In process III, valves 402 and 406 were closed, and valves 401 , 403 , and 405 were opened to apply a pressure of −60 kPa to deliver 900 μL of the lysate 113 from the lysis chamber 101 to the waste chamber 107 via the membrane chamber 108 in 2 minutes.

[0190] (iv) Process IV

[0191] In process IV, after all the dissolved product 113 is delivered to the membrane chamber 108, valves 403 and 405 are closed, and valves 402 and 406 are opened to apply a pressure of 60 kPa to deliver 500 μL of cleaning solution 111 from the cleaning solution chamber 105 to the dissolution chamber 101 via the membrane chamber 108 within 3 minutes.

[0192] (v) Process V

[0193] Process V shows a state in which the cleaning liquid 111 is being transported from the cleaning liquid chamber 105 to the dissolution chamber 101 via the membrane chamber 108 .

[0194] (vi) Process VI

[0195] Process VI represents a state after the entire amount or a predetermined amount of the cleaning solution 111 is transferred from the cleaning solution chamber 105 to the dissolution chamber 101 via the membrane chamber 108 .

[0196] By adopting the above-described operation flow, the dissolved product 113 and the cleaning liquid 111 can be continuously conveyed without trapping air between them.

[0197] In addition, Fig.12 The pressure and time applied in step III are not limited to the above values. In addition, a combination of time and pressure that can deliver more than 90% of the dissolved product 113 is required. Furthermore, the time and pressure required to deliver almost all of the dissolved product 113 from the dissolution chamber 101 to the purification membrane 102 can also be confirmed by experiments and set in the implementation procedure.

[0198] Example 3

[0199] refer to Fig.13 , Fig.14 and Fig.15 , Example 3 is described below.

[0200] <Configuration Example of the Purification System 301 Having Main Parts of the Flow Channel Chip 114>

[0201] Fig.13 1 is a diagram showing a configuration example of a purification system 500 including the main part of the flow channel chip 114 of Example 3, that is, the main part of the flow channel chip 114 when the sample lysing step 202 is completed or in progress.

[0202] The purification system 500 includes a dissolution chamber 101, a cleaning liquid chamber 105, a waste liquid 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 port 109 may not be one, but may be provided for each function. In addition, for example, dedicated ports may be provided for the flow paths 510 and 511, respectively. The dissolution chamber 101 holds a dissolution product 113 inside, and the cleaning liquid chamber 105 holds a cleaning liquid 111 inside.

[0203] like Fig.13 As shown, the flow path 506 extending from the discharge port of the dissolution chamber 101 is connected to one of the two inlet ports (upper part or top surface: it may not be the top surface) of the membrane chamber 108. The flow path 508 extending from the discharge port of the membrane chamber 108 is connected to the inlet port of the lower part (bottom surface: it may not be the bottom surface) of the waste liquid chamber 107. The flow path 507 extending from the discharge port of the cleaning liquid chamber 105 is connected to the other inlet port of the membrane chamber 108. The flow path 511 extending from the port 109 is connected to the inlet port of the cleaning liquid chamber 105. In addition, another flow path 510 extending from the port 109 is connected to the upper part (top surface: it may not be the top surface) of the waste liquid chamber 107. Furthermore, the flow path 509 is also connected to the upper part (top surface: it may not be the top surface) of the waste liquid chamber 107.

[0204] In addition, if Fig.13 As shown, valve 501 is provided in flow path 506, valve 502 is provided in flow path 507, valve 503 is provided in flow path 509, valve 504 is provided in flow path 510, and valve 505 is provided in flow path 511.

[0205] <Purification process>

[0206] Fig.14 It is a diagram for explaining purification processes I to VI of Example 3. Fig.15 is with Fig.14Flowchart corresponding to the purification process shown.

[0207] (i) Process I

[0208] Process I indicates a state during or immediately after dissolution in the sample dissolution step 202 (step 1501). At this time, valves 501, 502, 503, 504, and 505 are closed.

[0209] (ii) Process II

[0210] In process II, valves 501 and 504 are opened (step 1502) to apply a pressure of -60 kPa to deliver 900 μL of lysate 113 from the lysis chamber 101 to the waste chamber 107 via the membrane chamber 108 in 2 minutes (step 1503).

[0211] (iii) Process III and IV

[0212] In process III, after the dissolved product 113 is completely transported to the membrane chamber 108, the valve 504 is closed, and the valves 502 and 505 are opened (step 1504), and the cleaning liquid 111 is transported from the cleaning liquid chamber 105 to the flow path 506 via the membrane chamber 108 in 30 seconds by applying a pressure of 30 kPa (step 1505). At this time, the air existing 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 liquid 111. In addition, the space 108_2 on the outlet side of the membrane chamber 108 is filled with the dissolved product 113. In this way, it is important to discharge the air in the membrane chamber 108 to the flow path 506 in process IV to achieve continuous transportation without air being sandwiched between the dissolved product 113 and the cleaning liquid 111.

[0213] (iv) Process V

[0214] After the cleaning liquid starts to enter the flow path 506 , the valve 501 is closed and the valve 503 is opened (step 1506 ), and the cleaning liquid 111 is sent from the cleaning liquid chamber 105 to the waste liquid chamber 107 via the membrane chamber 108 (step 1507 ).

[0215] (v) Process VI

[0216] Process VI represents a state after the entire amount or a predetermined amount of the cleaning solution 111 is transferred from the cleaning solution chamber 105 to the waste solution chamber 107 via the membrane chamber 108 .

[0217] By adopting the above-described operation flow, the dissolved product 113 and the cleaning liquid 111 can be continuously conveyed without trapping air between them.

[0218] Example 4

[0219] refer to Fig.15 , Fig.16 and Fig.17 , Example 4 is described below.

[0220] <Configuration Example of the Purification System 600 Having Main Parts of the Flow Channel Chip 114>

[0221] Fig.16 1 is a diagram showing a configuration example of a purification system 600 including the main part of the flow channel chip 114 of Example 4, that is, the main part of the flow channel chip 114 when the sample lysis process 202 is completed or in progress.

[0222] The purification system 600 includes a dissolution chamber 101, a cleaning liquid chamber 105, a waste liquid chamber 107, a membrane chamber 108, valves 601, 602, 603, 604, 605, and 606, flow paths 607, 608, 609, 610, 611, 612, and 613 connecting the chambers, and a port 109. The port 109 may not be one, but may be provided for each function. For example, dedicated ports may be added to the flow paths 612 and 613, respectively. The dissolution chamber 101 holds a dissolved product 113 inside, and the cleaning liquid chamber 105 holds a cleaning liquid 111 inside.

[0223] like Fig.16 As shown, the flow path 606 extending from the discharge port of the dissolution chamber 101 and the flow path 608 extending from the discharge port of the cleaning liquid chamber 105 converge at a connection point (confluence point) 614 to form a flow path 609, and the flow path 609 is connected to the inlet of the upper part (top surface: it may not be the top surface) of the membrane chamber 108. The flow path 610 extending from the discharge port provided at the lower part (bottom surface: it may not be the bottom surface) of the membrane chamber 108 is connected to the inlet provided at the lower part (bottom surface: it may not be the bottom surface) of the waste liquid chamber 107. In addition, the flow path 611 extending from the discharge port provided at the upper part (top surface: it may not be the top surface) of the membrane chamber 108 is connected to the upper part (side surface: it may not be the side surface, it may be the top surface) of the waste liquid chamber 107. The flow path 613 extending from the port 109 is connected to the inlet of the cleaning liquid chamber 105. Another flow path 612 extending from the port 109 is connected to the upper portion (top surface: may not be the top surface) of the waste liquid chamber 107. Furthermore, the flow path 615 is also connected to the upper portion (top surface: may not be the top surface) of the waste liquid chamber 107.

[0224] In addition, if Fig.16As shown, valve 601 is provided in flow path 607, valve 602 is provided in flow path 608, valve 603 is provided in flow path 615, valve 604 is provided in flow path 611, valve 605 is provided in flow path 612, and valve 606 is provided in flow path 613.

[0225] <Purification process>

[0226] Fig.17 4 is a diagram for explaining purification processes I to V of Example 4. Fig.15 same.

[0227] (i) Process I

[0228] In process I, it shows the state in which the dissolution is in progress or just after the dissolution is completed in the sample dissolution step 202. At this time, valves 601, 602, 603, 604, 605 and 606 are closed.

[0229] (ii) Process II

[0230] In process II, valves 601 and 605 were opened to apply a pressure of -60 kPa to deliver 900 μL of lysate 113 from lysis chamber 101 to waste chamber 107 via membrane chamber 108 in 2 minutes.

[0231] (iii) Process III

[0232] In process III, after all the dissolved product 113 is transported to the membrane chamber 108, valves 601 and 605 are closed, and valves 602, 606, 604 and 603 are opened to apply a pressure of 30 kPa to transport the cleaning liquid 111 from the cleaning liquid chamber 105 via the membrane chamber 108 to the flow path 611 in 30 seconds.

[0233] (iv) Process IV and Process V

[0234] In process IV, after the cleaning liquid 111 enters the flow path 611, the valve 604 is closed, and the cleaning liquid 111 is transported from the cleaning liquid chamber 105 to the waste liquid chamber 107 via the membrane chamber 108. At this time, the air existing in the space 108_1 on the inlet side of the membrane chamber 108 moves to the waste liquid chamber 107 via the flow path 611, and the space 108_1 of the membrane chamber 108 is filled with the cleaning liquid 111. In addition, the space 108_2 on the outlet side of the membrane chamber 108 is filled with the dissolved product 113. In this way, in process IV, it is important to discharge the air in the membrane chamber 108 to the waste liquid chamber 107 via the flow path 611 to achieve continuous transportation without air being sandwiched between the dissolved product 113 and the cleaning liquid 111.

[0235] (v) Process VI

[0236] Process VI represents a state after the entire amount or a predetermined amount of the cleaning solution 111 is transferred from the cleaning solution chamber 105 to the waste solution chamber 107 via the membrane chamber 108 .

[0237] By adopting the above-described operation flow, the dissolved product 113 and the cleaning liquid 111 can be continuously conveyed without trapping air between them.

[0238] Example 5

[0239] Example 5 is a derivative form of Example 1. The purification system 301' of Example 5 is a derivative form of the purification system 301 of Example 1 (reference Figure 6 )The structure of valve 701 is added.

[0240] <Purification process>

[0241] Fig.18 It is a diagram for explaining purification processes I to V of Example 5.

[0242] (i) Process I

[0243] Process I indicates a state during or immediately after dissolution in the sample dissolution step 202. At this time, valves 302, 303, 304, 305, and 701 are closed.

[0244] (ii) Process II

[0245] In process II, valves 305, 304 and 701 were opened to apply a pressure of -60 kPa to deliver 900 μL of lysate 113 from lysis chamber 101 to waste chamber 107 via membrane chamber 108 in 1 minute.

[0246] (iii) Process III

[0247] In process III, before the air 702 following the transported dissolved product 113 exceeds the branch point (connection point (confluence point) 316 of the flow paths) between the flow paths 306 and 308 and the flow path 307, the valves 304 and 701 are closed, and the valves 302 and 303 are opened, so that the cleaning liquid 111 is transported from the cleaning liquid chamber 105 to the dissolution chamber 101 via the flow path 306 in 30 seconds by applying a pressure of 30 kPa. At this time, the air in the flow path 307 moves to the flow path 308 or 306, and the flow path 307 is filled with the cleaning liquid 111. Exhausting the air in the flow path 307 in process III is important for achieving continuous transport without air being caught between the dissolved product 113 and the cleaning liquid 111.

[0248] (iv) Process IV

[0249] In process IV, the valves 315 and 701 are opened, and 500 μL of the cleaning solution 111 is sent from the cleaning solution chamber 105 to the waste solution chamber 107 via the membrane chamber 108 at 60 kPa over 3 minutes.

[0250] (v) Process V

[0251] Process V represents a state after the entire amount or a predetermined amount of the cleaning solution 111 is transferred from the cleaning solution chamber 105 to the waste solution chamber 107 via the membrane chamber 108 .

[0252] By adopting the above-described operation flow, the dissolved product 113 and the cleaning liquid 111 can be continuously conveyed without trapping air between them.

[0253] Example 6

[0254] Example 6 is a derivative form of Example 2. The purification system 400' of Example 6 is a derivative form of the purification system 400 of Example 2 (refer to Fig.11 )The structure of valve 801 is added.

[0255] <Purification process>

[0256] Fig.19 It is a diagram for explaining purification processes I to V of Example 6.

[0257] (i) Process I

[0258] Process I indicates a state during or immediately after dissolution in the sample dissolution step 202. At this time, valves 401, 402, 403, 404, 405, 406, and 801 are closed.

[0259] (ii) Process II

[0260] In process II, valves 401 , 403 , 405 and 801 were opened to apply a pressure of −60 kPa to deliver 900 μL of lysate 113 from lysis chamber 101 to waste chamber 107 via membrane chamber 108 in 2 minutes.

[0261] (iii) Process III

[0262] In process III, when the dissolved product 113 is completely transferred to the membrane chamber 108, the valves 405 and 801 are closed, the valves 402, 406 and valve 404 are opened, and the cleaning liquid 111 is squeezed out from the cleaning liquid chamber 105 to the waste liquid chamber 107 at a pressure of 30 kPa or less in 30 seconds or less. At this time, the air in the flow path 409 moves to the flow path 410 or the waste liquid chamber 107. At this time, the air in the flow path 409 moves to the flow path 408 or 410, and the flow path 409 is filled with the cleaning liquid 111. In this way, exhausting the air in the flow path 409 in process III is important for achieving continuous transfer without air being trapped between the dissolved product 113 and the cleaning liquid 111.

[0263] (iv) Process IV

[0264] In process IV, the valve 403 is closed, the valve 801 is opened, and 500 μL of the cleaning solution 111 is sent from the cleaning solution chamber 105 to the dissolution chamber 101 via the membrane chamber 108 at 60 kPa over 3 minutes.

[0265] (v) Process V

[0266] Process V represents a state after the entire amount or a predetermined amount of the cleaning solution 111 is transferred from the cleaning solution chamber 105 to the dissolution chamber 101 via the membrane chamber 108 .

[0267] By adopting the above-described operation flow, the dissolved product 113 and the cleaning liquid 111 can be continuously conveyed without trapping air between them.

[0268] (3) Supplementary explanations on various embodiments

[0269] (i) In the case of Example 1, in order to continuously deliver the cleaning solution and the dissolved product, it is necessary to carry out the Figure 6 The switching from process III to process IV is shown. If the delivery of the dissolved product 113 is too long, air will enter the flow path 308, and if it is too short, a large amount of dissolved product 113 will remain in the dissolution chamber 101, and the yield will be reduced. Therefore, it is necessary to perform highly reproducible delivery or define the delivery time using a liquid level detection sensor.

[0270] However, when the sample types are various or the viscosity of the liquid is high, the reproducibility of the delivery of the dissolved product 113 becomes low. In addition, when a liquid level detection sensor is used, the components of the device increase, so the cost increases. In addition, there is also a risk that the liquid level detection sensor does not operate correctly.

[0271] On the other hand, the Fig.11 The switching from process III to process IV shown in Example 3 Fig.14The switching from process II to process III shown in the example 4 Fig.17 In the switching from process II to process III shown in the figure, even if the time required for conveying the dissolved product 113 is exceeded, the cleaning liquid 111 and the dissolved product 113 can be continuously conveyed. Therefore, in these embodiments, even without the liquid level detection sensor used in embodiment 1, the yield can be maintained, and a structure without the need for a liquid level detection sensor can be realized.

[0272] (ii) The lysate 113 contains polymers and particles derived from the sample, and precipitates derived from the mixture with the lysis buffer, etc. These are captured by the purification membrane when the lysate 113 is transported, which may lead to a decrease in the transport efficiency of the cleaning solution 111 and the eluent in the back end and a decrease in the transport efficiency of the fluid (air, nitrogen, or other gas) during drying.

[0273] In this regard, in the case of Example 3, since the cleaning liquid 111 flows from the side opposite to the dissolved product 113, there is also an effect of pushing the objects captured by the membrane when the dissolved product is transported, thereby realizing smooth transportation.

[0274] (iii) In the case of Example 3, Example 4, Example 5 (derivative form of Example 1) and Example 6 (derivative form of Example 2), it is not necessary to transport the cleaning solution 111 in multiple times. Therefore, it is not necessary to make the cleaning solution chamber 105 a structure that can cope with multiple transports. For example, as a reagent chamber, a reagent transport package using an actuator as shown in US2006-134773A1 can be used. However, in the case of Example 5 (derivative form of Example 1) and Example 6 (derivative form of Example 2), when valve 701 or valve 801 is not provided, there is a possibility that the fluid (air, nitrogen, other gases) moves to one side of the membrane chamber 108. Therefore, although it is possible to transport without valve 701 and valve 801, valve 701 or valve 801 can be provided to achieve more stable solution transport. In addition, regarding the flow path of Example 3, there is a tendency for the flow path to be longer than that of Example 1. Furthermore, regarding the flow path of Example 4, an additional flow path is required for Example 1.

[0275] (4) Summary of the Examples

[0276] In each embodiment, in a flow path with a purification membrane, it is not necessary to provide a dedicated structure for venting bubbles in the flow path chip, and two solutions can be continuously delivered to the portion where the membrane is provided, and the pressure required for delivering the solution can be reduced. As a result, the number of gas-liquid interfaces with high Laplace pressure can be reduced or eliminated, and the volume of the fluid (air, nitrogen, other gas) that must be delivered in a wet state of the membrane can be reduced or eliminated.

[0277] (i) According to Example 1 (reference Figure 5or 6), in the flow path of the purification system (biomolecule analysis device) 301 in which the membrane chamber 108 is arranged between the dissolution chamber (first chamber) 101 and the cleaning liquid chamber (second chamber) 105 and the waste liquid chamber 107, the cleaning liquid (second liquid) 111 is conveyed to a position at least exceeding the confluence point 316 of the flow paths 306 to 309 (first flow path) connecting the dissolution chamber 101 and the waste liquid chamber 107 and the flow path 307 (second flow path) extending from the cleaning liquid chamber (second chamber) 105, and the fluid (air, nitrogen, other gas) is discharged from the flow path 307 (second flow path). Then, the dissolved product (first liquid) 113 is conveyed from the dissolution chamber (first chamber) 101 to the waste liquid chamber 107 via the membrane chamber 108 under the control of the computer 115. Then, the cleaning liquid (second liquid) 111 in the cleaning liquid chamber (second chamber) 105 is transferred from the cleaning liquid chamber (second chamber) 105 to the waste liquid chamber 107 via the membrane chamber 108 under control of the computer 115. In this way, the dissolved product 113 and the cleaning liquid 111 can be continuously transferred to the waste liquid chamber 107 without mixing air between the dissolved product 113 and the cleaning liquid 111 during the solution transfer. Since the fluid (air, nitrogen, other gases) is not mixed, the pressure during the solution transfer can be reduced (refer to Fig.10 ), so even if the flow path is structurally fragile, it can be adopted, which can suppress the manufacturing cost of the purification system 301.

[0278] (ii) According to Example 2 (reference Fig.11 and 12), a membrane chamber 108 is arranged between the dissolution chamber (first chamber) 101 and the cleaning liquid chamber (second chamber) 105, and in the flow path of the purification system (biomolecule analysis device 400) in which the membrane chamber 108 and the cleaning liquid chamber 105 are connected to the waste liquid chamber 107, the cleaning liquid (second liquid) 111 is transported to a position at least exceeding the confluence point 414 of the flow paths 407 to 408 (first flow path) connecting the dissolution chamber (first chamber) 101 and the waste liquid chamber 107 and the flow path 409 (second flow path) extending from the cleaning liquid chamber (second chamber) 105, and the fluid (air, nitrogen, other gas) is discharged from the flow path 409 (second flow path). Then, the dissolved product (first liquid) 113 is transported from the dissolution chamber (first chamber) 101 to the waste liquid chamber 107 via the membrane chamber 108 under the control of the computer 115. Then, the cleaning liquid (second liquid) 111 of the cleaning liquid chamber (second chamber) 105 is transported from the cleaning liquid chamber (second chamber) 105 to the dissolution chamber (first chamber) 101 via the membrane chamber 108 under control of the computer 115. In this way, the cleaning liquid 111 is continuously transported to the dissolution chamber 101 after the dissolved product 113 is transported to the waste liquid chamber 107 without mixing the fluid (air, nitrogen, other gases) between the dissolved product 113 and the cleaning liquid 111 during the solution transport. Since the fluid (air, nitrogen, other gases) is not mixed, the pressure during the solution transport can be reduced (refer to the same as in Example 1). Fig.10 ), so even if the flow path is structurally fragile, it can be adopted, which can suppress the manufacturing cost of the purification system 400.

[0279] (iii) According to Example 3 (reference Fig.13 and Fig.12), in the flow path of the purification system (biomolecule analysis device) 500, the membrane chamber has a first inlet connected to the dissolution chamber (first chamber) 101 on the upstream side, a second inlet connected to the cleaning liquid chamber (second chamber) 105, and a discharge port connected to the waste liquid chamber 107, the flow path 506 (first flow path) extending from the dissolution chamber (first chamber) 101 is connected to the first inlet of the membrane chamber 108, the flow path 507 (second flow path) extending from the cleaning liquid chamber (second chamber) 105 is connected to the second inlet of the membrane chamber 108, and the flow path 508 (third flow path) extending from the discharge port of the membrane chamber 108 is connected to the waste liquid chamber 107, and the computer 115 is used to transport the dissolved product (first liquid) 113 from the dissolution chamber (first chamber) 101 to the waste liquid chamber 107 via the membrane chamber 108. Next, the cleaning liquid (second liquid) is fed from the cleaning liquid chamber (second chamber) 105 from 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 by the computer 115, and the cleaning liquid (second liquid) 111 is discharged from the first inlet of the membrane chamber 108 to the flow path 506 (first flow path) side in a state where the space 108_1 is filled with the cleaning liquid (second liquid) 111. Then, the cleaning liquid (second liquid) is fed from the cleaning liquid chamber (second chamber) to the waste liquid chamber 107 via the membrane chamber 108 by the computer 115 in a state where the cleaning liquid (second liquid) 111 is present on the flow path 506 (first flow path) side. In this way, the dissolved product 113 and the cleaning liquid 111 can be continuously fed to the waste liquid chamber 107 without mixing the fluid (air, nitrogen, or other gas) between the dissolved product 113 and the cleaning liquid 111 during the solution feeding. Since no fluid (air, nitrogen, other gases) is mixed in, the pressure during solution delivery can be reduced (refer to Example 1). Fig.10 ), so even if the flow path is structurally fragile, it can be adopted, which can suppress the manufacturing cost of the purification system 500.

[0280] (iv) According to Example 4 (reference Fig.16 and Fig.17), the flow path of the purification system (biomolecule analysis device) 600 includes: a membrane chamber 108 having a purification membrane 102, an inlet connected to the dissolution chamber (first chamber) 101 and the cleaning liquid chamber (second chamber) 105 via a flow path 609, a connection port to the waste liquid chamber 107, and a discharge port; flow paths 607 to 609 (first flow path) extending from the dissolution chamber (first chamber) 101 and connected to the inlet of the membrane chamber 108; and flow paths 607 to 609 (first flow path) extending from the cleaning liquid chamber (second chamber) 105 to the connection port connected to the flow paths 607 to 609. 09 (first flow path); a flow path 611 (connecting flow path) connecting the connection port of the first space 108_1 provided between the inlet of the membrane chamber 108 and the purification membrane to the waste liquid chamber 107; and a flow path 610 (third flow path) extending from the outlet of the membrane chamber 108 and connected to the waste liquid chamber 107, and the dissolved product (first liquid) 113 is transported from the dissolution chamber (first chamber) 101 to the waste liquid chamber 107 via the membrane chamber 108 by the computer 115. Next, the cleaning liquid (second liquid) 113 is transported from the cleaning liquid chamber (second chamber) 105 from the inlet of the membrane chamber 108 to the first space 108_1 and the flow path 611 (connecting flow path) by the computer 115. Then, the computer 115 is used to transfer the cleaning liquid (second liquid) 111 from the cleaning liquid chamber (second chamber) 105 to the waste liquid chamber 107 via the membrane chamber 108, while the first space 108_1 and the flow path 611 (connecting flow path) are filled with the cleaning liquid (second liquid) 111. In this way, the dissolved product 113 and the cleaning liquid 111 can be continuously transferred to the waste liquid chamber 107 without mixing the fluid (air, nitrogen, other gases) between the dissolved product 113 and the cleaning liquid 111 during the solution transfer. Since the fluid (air, nitrogen, other gases) is not mixed, the pressure during the solution transfer can be reduced (refer to the same as in Example 1). Fig.10 ), so even if the flow path is structurally fragile, it can be adopted, which can suppress the manufacturing cost of the purification system 600.

[0281] Description of Reference Numerals

[0282] 100 Biomolecule Analysis Device

[0283] 101 Dissolution Chamber

[0284] 102 Purification membrane

[0285] 103 Reaction Chamber

[0286] 104 Lysis buffer chamber

[0287] 105 cleaning fluid chamber

[0288] 106 Reagent Chamber

[0289] 107 Waste chamber

[0290] 108 membrane chamber

[0291] Port 109

[0292] 114 Flow Chip

[0293] 115 Computer

[0294] 301, 301', 400, 400', 500, 600 purification system

[0295] 313, 314 liquid level detection sensors.

Claims

1. A method for controlling liquid delivery in a flow path of a biomolecule analysis device using a computer, characterized in that: The biomolecule analysis device 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 the following processing: Controlled by the computer, the second liquid is delivered to a position at least beyond a confluence point of a first flow path connected from the first chamber to the waste liquid chamber and a second flow path extending from the second chamber, and a fluid different from the first liquid and the second liquid is discharged from the second flow path; Controlled by the computer, the first liquid is transported from the first chamber to the waste liquid chamber via the membrane chamber; and The computer is used to control the second liquid in the second chamber to be transported from the second chamber to the membrane chamber.

2. The method according to claim 1, characterized in that: The membrane chamber is arranged between the first chamber, the second chamber and the waste liquid chamber, The method further includes controlling, by the computer, to further transfer the second liquid delivered to the membrane chamber to the waste liquid chamber.

3. The method according to claim 1, characterized in that: The membrane chamber is arranged between the first chamber and the second chamber, The method further includes a process of controlling, by the computer, to further transfer the second liquid transferred to the membrane chamber to the first chamber.

4. The method according to claim 1, characterized in that: The computer controls the application of pressure and the opening and closing of valves in response to the input information of the flow rates of the first liquid and the second liquid, so as to perform the delivery of each of the first liquid and the second liquid.

5. The method according to claim 4, characterized in that: The membrane chamber is arranged between the first chamber, the second chamber and the waste liquid chamber, The computer (i) closes the first valve in the first flow path which is arranged on the side of the first chamber compared to the confluence point, and opens the second valve in the second flow path to apply infusion pressure, thereby transporting the second liquid to a position at least exceeding the confluence point, (ii) then closes the second valve and opens the first valve to apply infusion pressure, thereby transporting the first liquid from the first chamber to the waste liquid chamber via the membrane chamber, and (iii) further closes the first valve and opens the second valve again to apply infusion pressure, thereby transporting the second liquid from the second chamber to the waste liquid chamber via the membrane chamber.

6. The method according to claim 4, characterized in that: The membrane chamber is arranged between the first chamber and the second chamber, The computer (i) closes the first valve disposed between the first chamber and the membrane chamber, and opens the second valve disposed in the second flow path and the third valve disposed between the confluence point and the waste liquid chamber to apply infusion pressure, thereby delivering the second liquid to a position at least exceeding the confluence point, (ii) then closes the second valve and opens the first valve and the third valve to apply infusion pressure, thereby delivering the first liquid from the first chamber via the membrane chamber to the waste liquid chamber, and (iii) further closes the third valve and opens the first valve and the second valve to apply infusion pressure, thereby delivering the second liquid from the second chamber via the membrane chamber to the first chamber.

7. The method according to claim 4, characterized in that: The membrane chamber is arranged between the first chamber, the second chamber and the waste liquid chamber, The computer (i) opens the first valve arranged between the confluence point and the first chamber and the second valve arranged between the confluence point and the membrane chamber, and closes the third valve arranged in the second flow path to apply infusion pressure, thereby transporting the first liquid from the first chamber to the waste liquid chamber via the membrane chamber to fill the space from the third valve to the inlet of the waste liquid chamber with the first liquid, (ii) then closes the second valve and opens the first valve and the third valve to apply infusion pressure, thereby transporting the second liquid to a position at least exceeding the confluence point, and (iii) further closes the first and second valves and opens the third valve to apply infusion pressure, thereby transporting the second liquid from the second chamber to the waste liquid chamber via the membrane chamber.

8. The method according to claim 4, characterized in that: The membrane chamber is arranged 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 confluence point, and a third valve provided between the confluence point and the waste chamber, and closes a fourth valve provided in the second flow path to apply a liquid transfer pressure, thereby transferring the first liquid from the first chamber to the waste chamber via the membrane chamber; (ii) then transfers the first liquid to a position at least beyond the confluence point; (iii) then, while maintaining a state in which the space between the purification membrane and the first liquid discharge port of the membrane chamber and the flow path between the first liquid discharge port and the second valve is filled with the first liquid, closes the second valve, and opens the third valve and the fourth valve to apply a liquid transfer pressure, thereby transferring the second liquid from the second chamber to the waste chamber and discharging a fluid different from the first liquid and the second liquid contained in the second flow path to the waste chamber; and (iv) further closes the third valve and opens the first valve, the second valve, and the fourth valve to apply a liquid transfer pressure, thereby transferring the second liquid from the second chamber to the first chamber via the membrane chamber.

9. The method according to claim 1, characterized in that: The cross-sectional area of ​​the first flow path connected to the membrane chamber is 314 mm 2 the following.

10. The method according to claim 1, characterized in that: The purification membrane is capable of retaining particles larger than 0.1 μm.

11. The method according to claim 1, characterized in that: The purification membrane is composed of silicon dioxide.

12. The method according to claim 1, characterized in that: The first liquid contains: (i) a biopolymer including nucleic acids, proteins, lipids or polysaccharides, or a biomonomer including amino acids, lipids, sugars or nucleic acid bases; and (ii) a substance having molecules containing the biopolymer or derivatives of the biomonomer in its structure.

13. The method of claim 1, wherein: The second liquid has a faster evaporation rate, a lower surface tension, or a larger contact angle with the film than the first liquid.

14. The method of claim 1, wherein: The first liquid is a dissolved product, the second liquid is a cleaning liquid, and the fluid is air.

15. The method of claim 1, wherein: After the first liquid passes through the purification membrane, the computer transfers the first liquid and the second liquid at a pressure lower than a pressure required for the fluid to pass through the purification membrane.

16. The method of claim 4, wherein: The computer defines the switching timing of delivering the first liquid and the second liquid by using a liquid level detection sensor, or time, pressure or a combination thereof.

17. The method of claim 1, wherein: The volume of the second fluid is greater than the volume of the first space between the purification membrane and the inlet of the membrane chamber.

18. A method for controlling the liquid delivery in a flow path of a biomolecule analysis device by using a computer, characterized in that: The biomolecule analysis device 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, wherein the membrane chamber has a first inlet, a second inlet, and a discharge port, 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 discharge port of the membrane chamber is connected to the waste liquid chamber, The method comprises the following processing: Using the computer to transfer the first liquid from the first chamber to the waste chamber via the membrane chamber; Using the computer, the second liquid is transported from the second chamber to a first space between the first and second inlets of the membrane chamber and the purification membrane via the second inlet of the membrane chamber, and the second liquid is discharged from the first inlet of the membrane chamber to the first flow path in a state where the first space is filled with the second liquid; and Using the computer, the second liquid is transported from the second chamber to the waste liquid chamber via the membrane chamber in a state where the second liquid is present on one side of the first flow path.

19. The method of claim 18, wherein: The computer controls the application of pressure and the opening and closing of valves in response to the input information of the flow rates of the first liquid and the second liquid, so as to perform the delivery of each of the first liquid and the second liquid.

20. The method of claim 19, wherein: The computer (i) opens the first valve provided in the first flow path and closes the second valve provided in the second flow path to apply liquid infusion pressure, thereby transferring the first liquid from the first chamber to the waste liquid chamber via the membrane chamber; (ii) then, in a state where the second space between the purification membrane of the membrane chamber and the discharge port is filled with the first liquid, opens the first valve and the second valve to apply liquid infusion pressure, thereby filling the first space of the membrane chamber with the second liquid and discharging the second liquid from the first inlet port to the first flow path side; and (iii) further closes the first valve to apply liquid infusion pressure, thereby transferring the second liquid from the second chamber to the waste liquid chamber via the membrane chamber in a state where the second liquid is present on the first flow path side.

21. A method for controlling the liquid delivery in a flow path of a biomolecule analysis device by using a computer, characterized in that: The biomolecule analysis device 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 merges with the first flow path at a confluence point, and a connecting flow path is provided to connect the inlet of the membrane chamber to a first space between the purification membrane and the waste liquid chamber, The method comprises the following processing: Using the computer to transfer the first liquid from the first chamber to the waste chamber via the membrane chamber; Using the computer, transporting the second liquid from the second chamber to the first space and the connecting flow path via the inlet of the membrane chamber; and Using the computer, the second liquid is transferred from the second chamber to the waste liquid chamber via the membrane chamber in a state where the first space and the connecting flow path are filled with the second liquid.

22. The method of claim 21, wherein: The computer controls the application of pressure and the opening and closing of valves in response to the input information of the flow rates of the first liquid and the second liquid, so as to perform the delivery of each of the first liquid and the second liquid.

23. The method of claim 22, wherein: The computer: (i) opens the first valve provided in the first flow path and closes the second valve provided in the second flow path to apply liquid transfer pressure, thereby transferring the first liquid from the first chamber to the waste liquid chamber via the membrane chamber; (ii) then, in a state where the second space between the purification membrane and the discharge port of the membrane chamber is filled with the first liquid, closes the first valve and opens the second valve provided in the second flow path and the third valve provided in the connecting flow path to apply liquid transfer pressure, thereby filling the first space of the membrane chamber and the connecting flow path with the second liquid; (iii) further closes the third valve to apply liquid transfer pressure, thereby maintaining the second liquid in the connecting flow path and transferring the second liquid from the second chamber to the waste liquid chamber via the membrane chamber.

24. A biomolecule purification system, characterized in that include: A biomolecule analysis device having a flow path; and a computer for controlling the delivery of liquid 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; and a membrane chamber having a purification membrane, which is arranged between the first chamber, the second chamber and the waste liquid chamber, The computer performs the following processing: Controlling the second liquid to be delivered to a position at least beyond a confluence point of a first flow path connected from 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 liquid and the second liquid from the second flow path; Controlling the first liquid to be transported from the first chamber to the waste liquid chamber via the membrane chamber; and The second liquid in the second chamber is controlled to be transported from the second chamber to the waste liquid chamber via the membrane chamber.

25. A biomolecule purification system, characterized in that: include: A biomolecule analysis device having a flow path; and a computer for controlling the delivery of liquid 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, disposed between the first chamber and the second chamber; and Waste chamber, The computer executes: Controlling the second liquid to be delivered to a position at least beyond a confluence point of a first flow path connected from 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 liquid and the second liquid from the second flow path; Controlling the first liquid to be transported from the first chamber to the waste liquid chamber via the membrane chamber; and The process of controlling the second liquid in the second chamber to be transported from the second chamber to the first chamber via the membrane chamber.

26. A biomolecule purification system, characterized in that: include: A biomolecule analysis device having a flow path; and a computer for controlling the delivery of liquid 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 path extending from the first chamber and connected to the first inlet of the membrane chamber; A second flow path extending from the second chamber and connected to the second inlet of the membrane chamber; and A third flow path is extended from the discharge port of the membrane chamber and connected to the waste liquid chamber. The computer executes: a process of transferring the first liquid from the first chamber to the waste chamber via the membrane chamber; A process of transferring the second liquid from the second chamber to a first space between the first and second inlets of the membrane chamber and the purification membrane via the second inlet of the membrane chamber, and discharging the second liquid from the first inlet of the membrane chamber to the first flow path side in a state where the first space is filled with the second liquid; and A process of transferring the second liquid from the second chamber to the waste liquid chamber via the membrane chamber in a state where the second liquid exists on the first flow path side.

27. A biomolecule purification system, characterized in that: include: A biomolecule analysis device having a flow path; and a computer for controlling the delivery of liquid 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 connection port to the waste liquid chamber, and a discharge port; A first flow path extending from the first chamber and connected to the inlet of the membrane chamber; a second flow path extending from the second chamber to a confluence point with the first flow path; a connecting flow path connecting the connection port provided in the first space between the introduction port of the membrane chamber and the purification membrane with the waste liquid chamber; and The third flow path is extended from the discharge port of the membrane chamber and connected to the waste liquid chamber.

28. The biomolecule purification system of claim 27, wherein: The computer executes: a process of transferring the first liquid from the first chamber to the waste chamber via the membrane chamber; A process of transporting the second liquid from the second chamber to the first space and the connecting flow path via the inlet of the membrane chamber; and A process of transferring the second liquid from the second chamber to the waste liquid chamber via the membrane chamber in a state where the first space and the connecting flow path are filled with the second liquid.

Images