WO2005051524A1 - Procede et dispositif de separation et de recueil - Google Patents

Procede et dispositif de separation et de recueil Download PDF

Info

Publication number
WO2005051524A1
WO2005051524A1 PCT/JP2003/015264 JP0315264W WO2005051524A1 WO 2005051524 A1 WO2005051524 A1 WO 2005051524A1 JP 0315264 W JP0315264 W JP 0315264W WO 2005051524 A1 WO2005051524 A1 WO 2005051524A1
Authority
WO
WIPO (PCT)
Prior art keywords
carrier
target substance
liquid
substance
collecting
Prior art date
Application number
PCT/JP2003/015264
Other languages
English (en)
Japanese (ja)
Inventor
Akihiro Hino
Hideo Kuribara
Takashi Kodama
Yasutaka Minegishi
Shinji Kanayama
Satoshi Furui
Original Assignee
National Food Research Institute
Nippon Gene Co., Ltd.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by National Food Research Institute, Nippon Gene Co., Ltd. filed Critical National Food Research Institute
Priority to PCT/JP2003/015264 priority Critical patent/WO2005051524A1/fr
Priority to AU2003284488A priority patent/AU2003284488A1/en
Priority to JP2005510918A priority patent/JPWO2005051524A1/ja
Priority to JP2005515787A priority patent/JP5061288B2/ja
Priority to PCT/JP2004/017486 priority patent/WO2005052541A2/fr
Publication of WO2005051524A1 publication Critical patent/WO2005051524A1/fr

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/40—Concentrating samples
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/40—Concentrating samples
    • G01N1/405—Concentrating samples by adsorption or absorption
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
    • G01N1/31—Apparatus therefor
    • G01N2001/317—Apparatus therefor spraying liquids onto surfaces

Definitions

  • the present invention relates to a method and an apparatus for separating and collecting substances contained in a liquid such as a suspension or a solution.
  • deoxyliponucleic acid DNA
  • pickled salts such as guanidine thiosuccinate and sodium iodide.
  • an anion exchanger for example, see Japanese Patent Publication No. 28680462
  • the amount of lead-out is equal to the capacity of the syringe. Requires a lot of power to operate.
  • the washing operation and the like are repeatedly performed after being led out to the carrier, the It is inefficient because the syringe must be removed and the piston must be pulled out of the syringe that has been pushed in. Disclosure of the invention
  • an object of the present invention is to provide an inexpensive separation method in which a substance to be separated and collected contained in a liquid is efficiently brought into contact with a carrier having a property of separating from the liquid to perform separation, washing, and collection. It is an object of the present invention to obtain a collecting method and an inexpensive and compact separating and collecting apparatus.
  • a liquid containing the target substance is supplied to a carrier capable of collecting the target substance, the target substance is separated from the liquid, and held on the carrier.
  • the liquid is used at an injection pressure of a spray can to flow the liquid toward the carrier.
  • the liquid is caused to flow toward the carrier using the injection pressure of a spray can that is available at a low cost.
  • the target substance to which the present invention is applied and the liquid containing the target substance include the target substance which can be retained on the carrier when it comes into contact with the carrier, and the target substance in any form, such as dissolution or suspension.
  • Any liquid include biological substances such as sugars, proteins, and nucleic acids, and chemical substances such as metal ions and organic substances.
  • Examples of the liquid include solutions and suspensions containing one or more of these target substances.
  • the carrier capable of collecting the target substance referred to in the present invention may be any substance that can adsorb, bind or hold and separate the target substance when it comes into contact with a liquid containing the target substance. Includes those that select target substances by effect. Preferably, those having a chemical property of binding to the target substance, particularly those having a chemical property of selectively binding to the target substance are also suitable.
  • the shape of the carrier may be any shape as long as the liquid containing the target substance flows and comes into contact with the carrier so as to collect the target substance without escaping.
  • So-called "filter media” such as aggregates of sponge-like and porous particles provided with And a film-like or sponge-like material is particularly preferable.
  • the material of the carrier various materials such as paper and ceramic can be used, but those made of glass fiber or resin fiber are particularly preferable.
  • the spray can used in the present invention is a substance which becomes a gas at normal temperature and normal pressure, such as air, oxygen, carbon dioxide, or chlorofluorocarbon, and a mixture thereof is pressure-filled in a pressure-resistant can, and the injection port and the inside of the can are sealed. It refers to a device that has a communication path and opening and closing means for opening and closing the communication path.
  • the substance to be pressurized and filled into the pipe may be any substance that does not adversely affect the target substance. For example, oxygen, carbon dioxide, nitrogen or alternative fluorocarbon is preferable. Those used to blow off dust and the like attached to the contacts are commercially available at low cost.
  • the injection pressure generated from the spray can for the flow of the liquid containing the target substance, the above-described problems in the method using the centrifugal force or the method using the suction force are solved, and as a result, the target substance is collected.
  • Equipment can be used anywhere.
  • the injection pressure is sufficient to the extent permitted for commercially available spray cans, and is approximately 0.8 MPa or less.
  • any method may be used as long as the liquid containing the target substance is caused to flow toward the target substance collecting carrier by pressurizing with the injection pressure.
  • a carrier For example, by forming a carrier into a tube and circulating and flowing the liquid through this tubular carrier, unnecessary liquids or non-target substances contained in the liquid are eliminated, and the target substance inside the liquid is concentrated and separated.
  • the gas in the spray can is injected into the liquid to generate a flow that flows through the tubular carrier.
  • the inside of the pressure vessel is required.
  • a gas in a spray can is introduced into the vessel, and the liquid in the pressure-resistant container is extruded into a flow path by the spray pressure of the spray can and discharged toward the carrier.
  • the carrier that separates the target substance from the liquid is a liquid extruded with the spray pressure of a spray can by forming a pipe path using a carrier that has a chemical property that selectively binds to the target substance.
  • the target substance may be selectively chemically bonded while passing through the inside of the pipe route.
  • the liquid extruded at the injection pressure of a spray can may be passed through using a filter medium as a carrier.
  • the method of passing the liquid through the filter medium is preferable in the present invention using a spray can in which a limited volume of gas is sealed.
  • the present invention provides the following separation and collection method. That is, after supplying the liquid containing the target substance to a carrier capable of collecting the target substance, separating the target substance from the liquid and holding the target substance on the carrier, when collecting the target substance from the carrier, The liquid containing the target substance is introduced into the pressure vessel, connected to the filter material consisting of a carrier capable of collecting the target substance, and then connected to the gas injection port of the spray can to filter the liquid by the spray pressure of the spray can. At the same time as collecting the target substance on the filter medium, and discharging the liquid from which the target substance has been removed from the filter medium on which the target substance has been collected. Thereafter, an eluate for eluting the target substance was introduced into the pressure-resistant container, and connected to the filter medium that collected the target substance, and then the eluate was introduced into the filter medium and retained by the filter medium. Releases target substances.
  • the present invention provides a method for separating and collecting a substance comprising the following three steps.
  • a spray can Removably connect a spray can, a pressure-resistant container such as a syringe, and a filter medium made of a carrier capable of collecting the target substance.
  • a liquid containing the target substance is added into the pressure vessel. This is connected to the gas injection port of the spray can and the liquid is extruded to the filter medium by the spray pressure of the spray can, so that the target substance is trapped on the filter medium and the liquid from which the target substance has been removed is removed. Is discharged from the collected filter media.
  • the operation of pushing out the liquid containing the target substance to the filter medium may be performed not only once but also several times as necessary, in order to hold almost all of the target substance in the liquid on the filter medium.
  • the liquid containing the target substance may contain a reagent composition for promoting the binding of the target substance to a carrier (filter material) as necessary.
  • the eluate for eluting the target substance is introduced into the pressure vessel, and the eluate is introduced into the carrier (filter material) to collect the liquid containing the target substance released from the carrier. If the target substance has a size that cannot be eluted and pass from the carrier (filter material), the eluate may be flowed in the reverse flow to the method described in 1). That is, according to a specific preferred embodiment of the present invention, the method further comprises an operation of washing and removing components other than the target substance after holding the target substance on a carrier.
  • an operation of releasing the target substance from the carrier is performed by providing an eluate for eluting the target substance from the carrier holding the target substance by adsorption. It is further characterized by being provided.
  • the apparatus for separating and collecting a substance according to the present invention is an apparatus used for the above-described method for separating and collecting a substance, wherein a can container in which a substance that becomes a gas at normal temperature and normal pressure is filled therein, an injection port, A spray can provided with a path for communicating the injection port with the inside of the can, opening and closing means for opening and closing the path, a blowing port connectable to the injection port of the spray can, and a discharge port communicating with the outside.
  • a pressure-resistant container capable of storing a liquid therein; and a carrier detachably connected to the outlet and capable of collecting the target substance held in a container through which the liquid from the outlet can pass. It is provided with. This makes it possible to provide an inexpensive and compact device for performing adsorption, washing, separation and collection.
  • FIG. 1 is an explanatory view including a configuration and a partial cross section of a DNA collecting apparatus according to an embodiment of the present invention.
  • FIG. 2 is a schematic diagram showing the results of agarose gel electrophoresis. BEST MODE FOR CARRYING OUT THE INVENTION
  • the target substance and the target substance are captured.
  • Some principle is not important. Therefore, the target substances are not limited.
  • the gist of the present invention is that the implementation of the above-described principle is portable and can be embodied by the injection pressure of a disposable spray can.
  • FIG. 1 is an explanatory view including a configuration and a partial cross section of a DNA collecting apparatus according to an embodiment of the present invention.
  • the starting sample liquid used was a liquid obtained by adding 10 g of ADNA to 0.5 ml of a binding buffer containing guanidine hydrochloride, which is a chaotropic salt.
  • a syringe filter-GF / B (manufactured by Whatman) 13 in which a glass fiber sheath 11 was enclosed in a resin container 12 was used as a carrier capable of selectively binding DNA.
  • the inlet 14 of the syringe filter 13 was connected to the outlet 1.6 of a 1 ml syringe 15 (manufactured by Terumo Corporation) as a pressure-resistant container, and the liquid was added to the 1 ml syringe 15.
  • sprayer (trade name: OA dust cleaner, manufactured by Rix Co., Ltd.) By connecting the injection port 20 at the rear end of the syringe 15 to the injection port 19 of the head part 17 of 17 A device for collecting material was set up. After the connection was confirmed, the gas in the syringe can 17 was pressed by pressing the gas injection button 10 of the spray can 17, and the liquid in the syringe 15 was discharged from the syringe filter 13.
  • the buffer used had the following composition:
  • Binding buffer 8M guanidine hydrochloride
  • FIG. 2 is a schematic diagram showing the results of agarose gel electrophoresis.
  • lanes 21 and 24 are molecular weight markers
  • lane 22 is ADNA added to the binding buffer (before addition)
  • lane 23 is collected ADNA. As shown in Fig. 2, it was confirmed that the DNA to be collected was successfully collected.
  • the target substance could be achieved with only an extremely inexpensive apparatus as compared with the conventional method.
  • the place is not limited to the vicinity of the apparatus installation place, so that the target substance can be collected even in a place where there is no experimental facility.

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  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Sampling And Sample Adjustment (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Treatment Of Liquids With Adsorbents In General (AREA)
  • Extraction Or Liquid Replacement (AREA)

Abstract

L'invention concerne un procédé de séparation et de recueil comprenant les étapes consistant à alimenter des substances objectives renfermant du liquide dans un support capable de recueillir lesdites substances, aux fins de séparation de celles-ci du liquide et de maintenir les substances sur le support; et à recueillir les substances objectives à partir du support. Le liquide peut s'écouler en direction du support grâce à la pression d'injection d'un contenant pulvérisateur. Idéalement, le procédé comprend également une opération consistant à éliminer, par lavage, des substances qui ne sont pas les substances objectives, une fois celles-ci maintenues sur le support. La séparation, le lavage et le recueil des substances objectives peuvent être exécutés à faibles coûts, du fait que les substances séparées et recueillies peuvent venir en contact avec le support possédant des propriétés permettant de maintenir les substances objectives et les dimensions d'un dispositif de séparation et de recueil peuvent être réduites.
PCT/JP2003/015264 2003-11-28 2003-11-28 Procede et dispositif de separation et de recueil WO2005051524A1 (fr)

Priority Applications (5)

Application Number Priority Date Filing Date Title
PCT/JP2003/015264 WO2005051524A1 (fr) 2003-11-28 2003-11-28 Procede et dispositif de separation et de recueil
AU2003284488A AU2003284488A1 (en) 2003-11-28 2003-11-28 Method and device for separation and collection
JP2005510918A JPWO2005051524A1 (ja) 2003-11-28 2003-11-28 分離捕集方法及び装置
JP2005515787A JP5061288B2 (ja) 2003-11-28 2004-11-25 分離捕集方法及び装置
PCT/JP2004/017486 WO2005052541A2 (fr) 2003-11-28 2004-11-25 Procede et dispositif de separation et de collecte

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/JP2003/015264 WO2005051524A1 (fr) 2003-11-28 2003-11-28 Procede et dispositif de separation et de recueil

Publications (1)

Publication Number Publication Date
WO2005051524A1 true WO2005051524A1 (fr) 2005-06-09

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Family Applications (2)

Application Number Title Priority Date Filing Date
PCT/JP2003/015264 WO2005051524A1 (fr) 2003-11-28 2003-11-28 Procede et dispositif de separation et de recueil
PCT/JP2004/017486 WO2005052541A2 (fr) 2003-11-28 2004-11-25 Procede et dispositif de separation et de collecte

Family Applications After (1)

Application Number Title Priority Date Filing Date
PCT/JP2004/017486 WO2005052541A2 (fr) 2003-11-28 2004-11-25 Procede et dispositif de separation et de collecte

Country Status (3)

Country Link
JP (1) JPWO2005051524A1 (fr)
AU (1) AU2003284488A1 (fr)
WO (2) WO2005051524A1 (fr)

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH03143559A (ja) * 1989-10-30 1991-06-19 Toyota Motor Corp 溶接粉塵洗浄廃液の浄化再生方法
JPH06238265A (ja) * 1993-02-16 1994-08-30 Netsushii Kogyo Kk 汚水処理装置

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH02182260A (ja) * 1989-01-06 1990-07-16 Koken Co Ltd 骨形成活性を有する粒状骨補てつ材
JPH0376585A (ja) * 1989-08-18 1991-04-02 Ngk Insulators Ltd 微生物による生成物の製造方法

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH03143559A (ja) * 1989-10-30 1991-06-19 Toyota Motor Corp 溶接粉塵洗浄廃液の浄化再生方法
JPH06238265A (ja) * 1993-02-16 1994-08-30 Netsushii Kogyo Kk 汚水処理装置

Also Published As

Publication number Publication date
JPWO2005051524A1 (ja) 2007-08-09
AU2003284488A1 (en) 2005-06-17
WO2005052541A2 (fr) 2005-06-09
WO2005052541A3 (fr) 2005-08-04

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