WO2012133450A1 - ソフトマテリアルのマイクロアレイ作製方法 - Google Patents

ソフトマテリアルのマイクロアレイ作製方法 Download PDF

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WO2012133450A1
WO2012133450A1 PCT/JP2012/057996 JP2012057996W WO2012133450A1 WO 2012133450 A1 WO2012133450 A1 WO 2012133450A1 JP 2012057996 W JP2012057996 W JP 2012057996W WO 2012133450 A1 WO2012133450 A1 WO 2012133450A1
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Prior art keywords
microarray
substrate
soft material
layer
soft
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Ceased
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PCT/JP2012/057996
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English (en)
French (fr)
Japanese (ja)
Inventor
檜山　聡
香織 栗林
弘晃 尾上
昌治 竹内
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NTT Docomo Inc
University of Tokyo NUC
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NTT Docomo Inc
University of Tokyo NUC
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Priority to US13/814,588 priority Critical patent/US8999443B2/en
Priority to CN201280003445.4A priority patent/CN103180735B/zh
Priority to EP12763427.7A priority patent/EP2594941B1/en
Publication of WO2012133450A1 publication Critical patent/WO2012133450A1/ja
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B81—MICROSTRUCTURAL TECHNOLOGY
    • B81C—PROCESSES OR APPARATUS SPECIALLY ADAPTED FOR THE MANUFACTURE OR TREATMENT OF MICROSTRUCTURAL DEVICES OR SYSTEMS
    • B81C1/00—Manufacture or treatment of devices or systems in or on a substrate
    • B81C1/00015—Manufacture or treatment of devices or systems in or on a substrate for manufacturing microsystems
    • B81C1/00206—Processes for functionalising a surface, e.g. provide the surface with specific mechanical, chemical or biological properties
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B05—SPRAYING OR ATOMISING IN GENERAL; APPLYING FLUENT MATERIALS TO SURFACES, IN GENERAL
    • B05D—PROCESSES FOR APPLYING FLUENT MATERIALS TO SURFACES, IN GENERAL
    • B05D1/00—Processes for applying liquids or other fluent materials
    • B05D1/32—Processes for applying liquids or other fluent materials using means for protecting parts of a surface not to be coated, e.g. using stencils, resists
    • B05D1/322—Removable films used as masks
    • B05D1/325—Masking layer made of peelable film
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N37/00—Details not covered by any other group of this subclass
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00277—Apparatus
    • B01J2219/00279—Features relating to reactor vessels
    • B01J2219/00306—Reactor vessels in a multiple arrangement
    • B01J2219/00313—Reactor vessels in a multiple arrangement the reactor vessels being formed by arrays of wells in blocks
    • B01J2219/00315—Microtiter plates
    • B01J2219/00317—Microwell devices, i.e. having large numbers of wells
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00277—Apparatus
    • B01J2219/00497—Features relating to the solid phase supports
    • B01J2219/00527—Sheets
    • B01J2219/00529—DNA chips
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00583—Features relative to the processes being carried out
    • B01J2219/00603—Making arrays on substantially continuous surfaces
    • B01J2219/00605—Making arrays on substantially continuous surfaces the compounds being directly bound or immobilised to solid supports
    • B01J2219/00632—Introduction of reactive groups to the surface
    • B01J2219/00637—Introduction of reactive groups to the surface by coating it with another layer
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00583—Features relative to the processes being carried out
    • B01J2219/00603—Making arrays on substantially continuous surfaces
    • B01J2219/00659—Two-dimensional arrays
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00718—Type of compounds synthesised
    • B01J2219/0072—Organic compounds
    • B01J2219/00722—Nucleotides
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00718—Type of compounds synthesised
    • B01J2219/0072—Organic compounds
    • B01J2219/00725—Peptides
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B81—MICROSTRUCTURAL TECHNOLOGY
    • B81B—MICROSTRUCTURAL DEVICES OR SYSTEMS, e.g. MICROMECHANICAL DEVICES
    • B81B2201/00—Specific applications of microelectromechanical systems
    • B81B2201/02—Sensors
    • B81B2201/0214—Biosensors; Chemical sensors
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B81—MICROSTRUCTURAL TECHNOLOGY
    • B81B—MICROSTRUCTURAL DEVICES OR SYSTEMS, e.g. MICROMECHANICAL DEVICES
    • B81B2201/00—Specific applications of microelectromechanical systems
    • B81B2201/05—Microfluidics
    • B81B2201/058—Microfluidics not provided for in B81B2201/051 - B81B2201/054

Definitions

  • the present invention relates to a method for producing a microarray of soft material.
  • the present invention relates to a method for producing a microarray of a plurality of types of soft materials on the same substrate.
  • soft material may be derived from living organisms or non-living organisms, and polymers, gels, rubbers, colloids, micelles, liquid crystals, and various types in which small molecules are connected (or assembled) in various forms.
  • biopolymers nucleic acids such as DNA and RNA, proteins, carbohydrates, glycoproteins, etc.
  • Non-Patent Document 1 discloses a technique for forming a single type of soft material on a substrate in a general-purpose and scalable manner, but a technique for arraying different types of soft materials on the same substrate is disclosed. Not disclosed.
  • Non-Patent Document 2 discloses that a micropattern is formed on a polyparaxylylene resin vacuum-deposited on a substrate to form a stencil, and a plurality of types of soft materials are ejected onto the stencil in an ink jet printing manner to produce polyparaxylylene.
  • a technology is disclosed in which a lenic resin is peeled off from a substrate to produce a plurality of types of soft material microarrays on the same substrate.
  • this technique not only requires a high-performance spotter capable of discharging a soft material as an additional dedicated device, but also has a problem that the spot interval depends on the performance of the spotter. That is, depending on the performance of the spotter, in order to avoid contamination (unintentional mixing) between different types of soft materials, there is a problem that the spot interval for arraying different types of soft materials is limited.
  • Non-Patent Document 3 a micropattern is formed on a two-layer polyparaxylylene resin vacuum-deposited on a substrate to form a stencil, and a soft material is injected on the stencil to form a two-layer polyparaxylylene.
  • a technique is disclosed in which a plurality of different types of soft material microarrays are fabricated on the same substrate by peeling the resin one by one from the substrate one after another.
  • this technology has a problem that it cannot exclude the possibility of contamination between different types of soft materials.
  • Non-Patent Document 4 a micropattern is formed on a single layer of polyparaxylylene resin vacuum-deposited on a substrate to form a stencil, and a plurality of microchannels are formed on the stencil,
  • a technique for producing a microarray of a plurality of types of soft materials on the same substrate by injecting different types of soft materials and peeling the polyparaxylylene resin from the substrate.
  • this technology is unlikely to cause contamination, it is a method of connecting a micro flow path to each spot of the microarray, so the spot interval for arraying different types of soft materials cannot be set to a desired size, and is larger. There is a problem that it is difficult to produce an array of areas.
  • the present invention is a method for producing a soft material small-interval microarray using a polyparaxylylene resin lift-off method, which is versatile and scalable without contamination of multiple types of soft materials on the same substrate.
  • Another object of the present invention is to provide a method for producing a microarray of a plurality of types of soft materials.
  • the present inventor has obtained (i) (for example, a polyparaxylylene resin lift-off method).
  • the obtained soft material on the substrate is dehydrated without being denatured (by lyophilization etc.), so that the normal polyparaxylylene resin lift-off method can be applied to the substrate, and (ii) the soft material
  • the present inventors have found that the polyparaxylene resin deposited thereon can be smoothly peeled off from the substrate without impairing the fixing state, structure, and function of the soft material.
  • the present invention is a method for producing a microarray of a plurality of types of soft materials, a first layer of polyparaxylylene resin is vapor-deposited on a substrate, and a first micropattern is formed on the polyparaxylylene resin.
  • a first soft material solution is introduced to form a first microarray on the substrate surface, and the first soft material is lyophilized to form a first soft material microarrayed substrate.
  • the second layer of polyparaxylylene resin is deposited on the microarrayed substrate of the first soft material and laminated, and the first layer and the second layer of polyparaxylylene resin are penetrated.
  • a plurality of types of soft materials can be used on a single substrate for general purpose and scalable, without using a spotter, and with a small interval microarray. Can be produced.
  • the microarray manufacturing procedure according to the present invention not only two types of soft materials but also two or more types of soft materials can be manufactured.
  • the microarray substrate on which the lyophilized soft material is immobilized can be stored and delivered for a long time at room temperature or low temperature, and can be easily used simply by introducing a buffer solution when the user uses the substrate. be able to.
  • FIG. 1 is a diagram conceptually showing a method according to an embodiment of the present invention.
  • a method for producing a microarray of a plurality of types of soft materials according to an embodiment of the present invention uses a so-called conventional polyparaxylylene resin lift-off method, and a polyparaxylylene resin known so far Includes the lift-off method or its various applications.
  • the method according to the embodiment of the present invention is not particularly limited with respect to conditions, materials, apparatuses, reaction conditions, and the like of a normal polyparaxylylene resin lift-off method.
  • a first layer of polyparaxylylene resin is deposited on a substrate, and the first layer of polyparaxylylene resin is subjected to a first process.
  • Such a process is a process of applying a conventional polyparaxylylene resin lift-off method, and the obtained substrate has the first microarray formed (immobilized) thereon.
  • shape etc. There is no restriction
  • the first soft material introduced is usually fixed on the substrate in the form of an aqueous solution or an appropriate solution.
  • the soft material is fixed to the board
  • the substrate surface is not particularly required to be modified to promote the binding with the soft material, but in some cases, a treatment for strengthening the binding with the soft material may be performed.
  • the method according to the embodiment of the present invention includes the step of lyophilizing the obtained first soft material to obtain a microarrayed substrate of the first freeze-dried soft material. .
  • the first soft material fixed to the substrate is freeze-dried.
  • “freeze-drying” is widely used as “from the soft material on the substrate. , Removing moisture and drying so that the structure and function of the soft material are not irreversibly denatured ”. Accordingly, the presence or absence of cooling, the presence or absence of a freezing buffer such as sucrose or trehalose, the presence or absence of a vacuum operation, etc. can be appropriately selected depending on the soft material on the lyophilized substrate.
  • the “freeze drying” can be performed on all or a part of the soft material on the substrate, or on the entire substrate.
  • the second-layer polyparaxylylene resin is again deposited on the substrate. Then, a second micropattern is formed on the second-layer polyparaxylylene resin.
  • the polyparaxylylene resin vapor deposition of the second layer is carried out including the surface of the first soft material fixed on the substrate and freeze-dried. Such vapor deposition is possible under the normal conditions of polyparaxylylene resin vapor deposition, and will be described below. Moreover, no damage such as modification is observed not only on the surface of the first soft material but also inside the vapor deposition. This is considered to be because the first soft material fixed on the substrate is sufficiently dried.
  • the condition of the normal polyparaxylylene resin lift-off method already described above can be applied as it is to form the second micropattern on the second-layer polyparaxylylene resin.
  • the second micropattern formed on the polyparaxylylene resin of the second layer is provided with the first and second soft material microarrays on the same surface of the substrate.
  • the second micropattern is formed on a substrate at a location different from the first micropattern by penetrating the second layer of polyparaxylylene resin.
  • the method according to the embodiment of the present invention further introduces a solution containing the second soft material into the second micropattern of the obtained second-layer polyparaxylylene resin, so that the second on the substrate.
  • the microarray is formed (immobilized). This step may be the same method as the introduction of the solution containing the first soft material above.
  • a polyparaxylylene resin is obtained from the substrate on which the obtained second microarray is formed (immobilized).
  • the microarray of the first and second soft materials can be manufactured on the same surface of the substrate.
  • the first soft material is in a dry state
  • the second soft material is in a solution (wet) state. If necessary, the whole is freeze-dried again to make the first and second soft materials dry, or a buffer solution or the like is injected to make the first and second soft materials wet. be able to. By freeze-drying the whole, it becomes possible to store at room temperature or low temperature.
  • the first polyparaxylylene resin is deposited on the substrate on which the first soft material is fixed and freeze-dried as a target for the second-layer polyparaxylylene resin deposition.
  • the embodiment of the present invention is not limited to this.
  • the first polyparaxylylene layer peeled off is freeze-dried, and the first soft material is introduced and fixed to the substrate.
  • the target is also freeze-dried and then peeled off the first polyparaxylylene layer.
  • a first layer of polyparaxylylene resin is vacuum-deposited on a substrate (FIG. 2A (a)).
  • the substrate may be a material that can fix a soft material and can withstand polyparaxylylene vapor deposition conditions. Examples thereof include glass, silicon, and plastic. When using a microarray, if it is preferable to be light-transmitting for the purpose of measurement or the like, glass that is light-transmitting is preferable.
  • the method for fixing the soft material to the surface of the substrate is not particularly limited. However, if necessary, the substrate surface can be subjected to a treatment for promoting the fixation in order to obtain stronger fixation.
  • the type of polyparaxylylene resin to be deposited is not particularly limited, but here, explanation will be made by taking parylene C (Japan Parylene Godo Kaisha) as an example. Other types of parylene N, parylene D, and parylene HT can also be selected as appropriate. (Both are available at Japan Parylene LLC).
  • the vapor deposition thickness of the polyparaxylylene resin is not particularly limited as long as a pattern having a structure in which a soft material is sufficiently introduced in a later step is formed. Specifically, it is desirable if it is 2 ⁇ m or less. Here, the description will be made by taking the case of vapor deposition with a thickness of 1 ⁇ m as an example.
  • a photoresist is laminated on the deposited polyparaxylylene resin (FIG. 2A (b)).
  • S1818 Silicone Far East Ltd.
  • the lamination method is not particularly limited, and an ordinary method such as spin coating is preferable.
  • photolithography is performed by ultraviolet irradiation through a glass photomask on which the first micropattern is drawn (FIG. 2A (c)), and further development of the photoresist is performed (FIG. 2A (d)).
  • the developer can be selected in combination with the photoresist used. For example, when S1818 is used as the photoresist, NMD-3 (available from Tokyo Ohka Kogyo Co., Ltd.) can be used as the developer.
  • dry etching with oxygen plasma is performed to draw a first micro pattern on the polyparaxylylene resin (FIG. 2A (e)).
  • photolithography by ultraviolet irradiation and dry etching by oxygen plasma have been described as examples.
  • the present invention is not limited to this, and may be lithography by an electron beam, for example.
  • a flow cell is formed on the micro-patterned first-layer polyparaxylylene resin (FIG. 2A (g)).
  • the flow cell is manufactured by using, for example, a silicon rubber sheet as a spacer and sandwiching it with a cover glass.
  • a solution containing a first soft material for which a microarray is to be produced is injected into the flow cell, and the first microarray is introduced and formed on the exposed glass substrate surface.
  • biotinylated BSA bovine serum albumin
  • 1 mg / mL streptavidin 1 mg / mL streptavidin
  • 5 ⁇ M biotinylated single-stranded DNA 23 base length, fluorescent dye FITC (fluorescein isothiocyanate) modification
  • the solution in the flow cell is replaced with a salt-free solution such as ultrapure water or distilled water.
  • a salt-free solution such as ultrapure water or distilled water.
  • the entire substrate including the flow cell is freeze-dried (as it is) using a freeze-drying apparatus (FIG. 2A (h)). Furthermore, after the solution containing the first soft material is completely dried, the flow cell is disassembled (FIG. 2A (h ′)). Here, the whole substrate not including the flow cell may be freeze-dried after the flow cell is disassembled.
  • a second layer of polyparaxylylene resin is vacuum-deposited and laminated to a thickness of, for example, 1 ⁇ m on the freeze-dried substrate under the same conditions and procedures as the first layer (FIG. 2B (i).
  • a photoresist such as S1818 is spin-coated and laminated (FIG. 2B (j)).
  • the baking condition of the photoresist may be a high temperature for a short time (for example, 100 ° C. for 1 minute) or a low temperature in consideration of the properties (for example, heat sensitivity) of the first lyophilized soft material.
  • a long time for example, 3 hours at 35 ° C. or conditions for combining them can be selected.
  • photolithography is performed by ultraviolet irradiation through a glass photomask on which a second micropattern is drawn (FIG. 2B (k)), and photoresist development is performed using NMD-3 (FIG. 2B (l)). .
  • the dry etching conditions for penetrating the first and second polyparaxylylene resins are not particularly limited, depending on the thickness of the first and second polyparaxylylene resins, for example, It can be easily selected by adjusting the etching time.
  • a solution containing the second soft material for which a microarray is to be produced is injected into the flow cell, and the second microarray is formed on the exposed glass substrate surface.
  • the second soft material 4 mg / mL biotinylated BSA, 1 mg / mL streptavidin, 5 ⁇ M biotinylated single-stranded DNA (23 base length, modified with fluorescent dye TMR (tetramethylrhodamine)) is the second soft material.
  • TMR fluorescent dye
  • the polyparaxylylene resin of the first layer and the second layer is peeled off simultaneously (FIG. 2B (p)).
  • the polyparaxylylene resin of the first layer and the second layer is easily peeled off at the same time by peeling off a part thereof with tweezers, for example.
  • FIG. 3 shows an example of the result of observation of the microarray produced by the above procedure with a fluorescence microscope.
  • FIG. (I) First, two different soft materials are fixed on the glass substrate to a sufficient extent for the following various treatments.
  • two different types of soft materials (labeled proteins constructed by combining the functions of a plurality of proteins, respectively) should function without being denatured.
  • two different types of soft materials (labeled proteins constructed by combining the functions of a plurality of proteins, respectively) should function without being denatured.
  • the FITC-modified single-stranded DNA as the first soft material and the TMR-modified single-stranded DNA as the second soft material are microarrayed on the same substrate without contamination. .
  • the arraying is microscale and the spacing between the spots forming the array is sufficiently well separated.
  • FIG. 4 is a fluorescence microscope observation image showing a result of an experiment for verifying whether the microarrayed single-stranded DNA (soft material) according to the embodiment of the present invention has the ability to form double strands. is there.
  • FIG. 4 confirms that the single-stranded DNA microarrayed by the method according to the embodiment of the present invention that has undergone the freeze-drying process retains its ability to form double strands, and is subjected to denaturation and the like. I understand that it is not. That is, the single-stranded DNA of the base sequence A that has undergone the freeze-drying process
  • the complementary strand specifically binds only to the spot where the single-stranded DNA of the base sequence A is immobilized (FIG. 4c), and the ability to form a double-stranded single-stranded DNA after lyophilization. was confirmed to be retained.
  • FIG. 5 is a fluorescence microscope observation image showing a result of a verification experiment on whether the microarrayed protein according to the embodiment of the present invention retains its functionality.
  • kinesin which is a motor protein
  • kinesin which is a motor protein
  • a kinesin microarray was prepared by peeling off the polyparaxylylene resin. The surface of the glass substrate outside the kinesin spot was coated with 5 mg / mL casein to suppress nonspecific adsorption.
  • microtubules labeled with the fluorescent dye TMR were introduced, the microtubules landed specifically only at spots where kinesin was immobilized, and microarrays of microtubules were formed (FIG. 5a).
  • the soft material (kinesin) obtained by performing the freeze-drying process in the manufacturing process according to the embodiment of the present invention retains the ability to bind to microtubules, and also slides the microtubules on a glass substrate. It has the activity to make it.
  • the microarray manufacturing method according to the embodiment of the present invention can be applied to DNA and proteins.
  • the embodiment of the present invention is not limited to DNA and protein, and any soft material such as RNA (ribonucleic acid), lipid, and microbeads can be used.
  • the manufacturing method according to the embodiment of the present invention includes a step of freeze-drying the soft material, this does not exclude application to the soft material in which the embodiment of the present invention is difficult to freeze-dry.
  • Some soft materials such as liposomes and cells that are lipid bilayer capsules, are not suitable for lyophilization, but by chemically modifying these surfaces with single-stranded DNA, biotin, antibodies, etc., It is also possible to produce a microarray. Methods for modifying DNA on liposomes and cell surfaces are disclosed, for example, in References 1 and 2 below, methods for biotinylation modification in Reference 3 and methods for antibody modification in Reference 4, respectively.

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PCT/JP2012/057996 2011-03-28 2012-03-27 ソフトマテリアルのマイクロアレイ作製方法 Ceased WO2012133450A1 (ja)

Priority Applications (3)

Application Number Priority Date Filing Date Title
US13/814,588 US8999443B2 (en) 2011-03-28 2012-03-27 Method for fabricating a microarray of soft materials
CN201280003445.4A CN103180735B (zh) 2011-03-28 2012-03-27 软物质的微阵列制作方法
EP12763427.7A EP2594941B1 (en) 2011-03-28 2012-03-27 Method for fabricating microarrays of soft materials

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JP2011-070934 2011-03-28
JP2011070934A JP5656192B2 (ja) 2011-03-28 2011-03-28 ソフトマテリアルのマイクロアレイ作製方法

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Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2010531972A (ja) * 2007-03-26 2010-09-30 ナノキシス アーベー 制御された単分子層形成のための方法及びデバイス

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6559474B1 (en) * 2000-09-18 2003-05-06 Cornell Research Foundation, Inc, Method for topographical patterning of materials
JP4007269B2 (ja) * 2003-07-29 2007-11-14 オムロン株式会社 生体分子の形状転写方法、チップ基板の製造方法及びバイオチップの製造方法
JP2006114489A (ja) 2004-09-14 2006-04-27 Nagoya Institute Of Technology 誘電体メタマテリアル、および磁性体メタマテリアル
JP2008014352A (ja) 2006-07-04 2008-01-24 Maezawa Kyuso Industries Co Ltd 逆止弁ユニットと濾過整流板
WO2008097360A2 (en) * 2006-09-14 2008-08-14 Board Of Supervisors Of Louisiana State University And Agricultural And Mechanical College Polymeric nanopillars and nanotubes, their manufacture and uses

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2010531972A (ja) * 2007-03-26 2010-09-30 ナノキシス アーベー 制御された単分子層形成のための方法及びデバイス

Non-Patent Citations (13)

* Cited by examiner, † Cited by third party
Title
A. ITO ET AL.: "Magnetite nanoparticle-loaded anti-HER2 immuno-liposomes for combination of antibody therapy with hyperthermia", CANCER LETTERS, vol. 212, 2004, pages 167 - 175
B. ILIC ET AL.: "Topographical Patterning of Chemically Sensitive Biological Materials Using a Polymer-Based Dry Lift Off", BIOMEDICAL MICRODEVICES, vol. 2, 2000, pages 317 - 322, XP055083830, DOI: doi:10.1023/A:1009911407093
B. ILIC ET AL.: "Topographical Patterning of Chemically Sensitive Biological Materials Using a Polymer-Based Dry Lift Off", BIOMEDICAL MICRODEVICES, vol. 2, no. 4, 2000, pages 317 - 322, XP055083830 *
C. P. TAN ET AL.: "Nanoscale Resolution, Multicomponent Biomolecular Arrays Generated by Aligned Printing with Parylene Peel-Off", NANO LETTERS, vol. 10, 2010, pages 719 - 725, XP055083833, DOI: doi:10.1021/nl903968s
C.P.TAN ET AL.: "Nanoscale Resolution, Multicomponent Biomolecular Arrays Generated By Aligned Printing With Parylene Peel-Off", NANO LETT., vol. 10, 2010, pages 719 - 725, XP055083833 *
D. STAMOU ET AL.: "Self-assembled microarrays of attoliter molecular vessels", ANGEWANDTE CHEMIE INTERNATIONAL EDITION, vol. 42, 2003, pages 5580 - 5583, XP001197495, DOI: doi:10.1002/anie.200351866
K. ATSUTA ET AL.: "A Parylene Lift-Off Process with Microfluidic Channels for Selective Protein Patterning", JOURNAL OF MICROMECHANICS AND MICROENGINEERING, vol. 17, 2007, pages 496 - 500
K. ATSUTA ET AL.: "A Parylene Lift-off Process with Microfluidic Channels for Selective Protein Patterning", JOURNAL OF MICROMECHANICS AND MICROENGINEERING, vol. 17, 2007, pages 496 - 500, XP020120026 *
K. KURIBAYASHI ET AL.: "Sequential Parylene Lift-Off Process for Selective Patterning of Biological Materials", PROC. IEEE MEMS'07, 2007, pages 501 - 504, XP031203868, DOI: doi:10.1109/MEMSYS.2007.4433114
KURIBAYASHI K. ET AL.: "Sequential Parylene Lift-off Process for Selective Patterning of Biological Materials", MEMS 2007, 2007, KOBE JAPAN, pages 501 - 504, XP031203868 *
S. C. HSIAO ET AL.: "Direct cell surface modification with DNA for the capture of primary cells and the investigation of myotube formation on defined patterns", LANGMUIR, vol. 25, 2009, pages 6985 - 6991, XP055063345, DOI: doi:10.1021/la900150n
S. HIYAMA ET AL.: "Biomolecular-motor-based autonomous delivery of lipid vesicles as nano- or microscale reactors on a chip", LAB ON A CHIP, vol. 10, 2010, pages 2741 - 2748
See also references of EP2594941A4

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JP5656192B2 (ja) 2015-01-21
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