JP4039297B2 - Extraction method and extraction device for biopolymer fixed to magnetic beads - Google Patents
Extraction method and extraction device for biopolymer fixed to magnetic beads Download PDFInfo
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- JP4039297B2 JP4039297B2 JP2003103714A JP2003103714A JP4039297B2 JP 4039297 B2 JP4039297 B2 JP 4039297B2 JP 2003103714 A JP2003103714 A JP 2003103714A JP 2003103714 A JP2003103714 A JP 2003103714A JP 4039297 B2 JP4039297 B2 JP 4039297B2
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- 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
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
- G01N33/54326—Magnetic particles
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- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
- C12N15/1006—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers
- C12N15/101—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers by chromatography, e.g. electrophoresis, ion-exchange, reverse phase
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- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
- C12N15/1006—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers
- C12N15/1013—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers by using magnetic beads
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- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
- C12N15/1017—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by filtration, e.g. using filters, frits, membranes
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- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
- G01N27/447—Systems using electrophoresis
- G01N27/44756—Apparatus specially adapted therefor
- G01N27/44786—Apparatus specially adapted therefor of the magneto-electrophoresis type
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- 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
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
- G01N33/559—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody through a gel, e.g. Ouchterlony technique
Description
【０００１】
【発明の属する技術分野】
本発明は、生体試料の細胞から生体高分子を抽出分離する抽出装置や、その前処理装置、あるいは細胞からの生体高分子の抽出、増幅、検出などを一体的に行うカートリッジなどにおいて使用されるもので、生体試料である生体高分子の中からターゲット生体高分子を分離して抽出する方法および装置に関するものである。
【０００２】
【従来の技術】
以下、生体高分子としてDNAを例にとって説明する。DNAチップに用いるターゲットDNAを生体細胞から抽出し回収する方法としては、大別して２種類の方法がある。１つは遠心分離による方法であり、他方は磁気ビーズによる方法である。
遠心分離による方法はそのための装置が大がかりであるため、小型化が望まれる将来の方向としては磁気ビーズによる方法が主流になると予測される（磁気ビーズの応用例としては例えば非特許文献１参照）。
【０００３】
磁気ビーズによる方法とは例えば次のような方法である。磁気ビーズの表面に、ある密度でプローブDNAあるいはプローブ抗体を固定し、溶液中のターゲットDNAとプローブとの相補結合により溶液中のDNAの回収を行い、その後磁石により磁気ビーズを集め、洗浄した後磁気ビーズ表面からDNAを溶液により解離し回収する方法である。
【０００４】
さて、このような磁気ビーズ方式を採用した装置として現在は卓上型パーソナルコンピュータ程度の大きさのものが実現されているが、その装置がチップ化され小型化されたものは未だ出現していない。
ただし、小型チップ化するために、μTAS（micro/miniaturized total analysis system）を活用したデバイスは、現在色々な分野で紹介されている（μTASについては例えば非特許文献２参照）。
【０００５】
【非特許文献１】
松永是監修、「DNAチップ応用技術」、株式会社シーエムシー、２０００年７月発行、第７章 磁気ビーズ利用DNAチップ 竹山春子，中山秀喜
【非特許文献２】
庄子 習一、「生化学 マイクロ化学分析システム−マイクロマシン技術−」、第１頁、第１および第２項、図１、［2003/02/26検索］、インターネット<URL:http://www.jaclap.org/LabCP/p11.html>
【０００６】
【発明が解決しようとする課題】
しかしながら、このμTASデバイスにおいて、駆動装置であるポンプや、制御装置であるバルブ、そして攪拌装置であるミキサーなどは実用に耐えるものがなかなか実現されておらず、そのため流体の移動を伴うμTASデバイスでは商品化されたものは多くない。
【０００７】
これは、ミクロの世界では流体の粘性や流路形状等により流体動特性が大きく変わり、経済的および機能的に課題を解決できる要素技術が未だ試行錯誤の段階にあるためと考えられる。
それゆえ、流体の移動を伴うことなく、磁気ビーズに固定のターゲット生体高分子を他の生体高分子から分離し抽出することのできる方式の出現が望まれている。
【０００８】
本発明の目的は、上記の課題を解決するもので、流体の移動を伴うことなく、磁気ビーズに固定のターゲット生体高分子を他の生体高分子から分離し抽出（以下単に抽出という）することができ、かつ小型化も図り得る磁気ビーズに固定の生体高分子の抽出方法および装置を提供することにある。
【０００９】
【課題を解決するための手段】
このような目的を達成するために、請求項１の方法の発明では、
生体試料から負の電荷を帯び磁気ビーズに固定されたターゲット生体高分子を抽出する方法であって、
前記生体試料を含む第１の溶液と、移動された生体高分子を保存するための第２の溶液と、抽出された磁気ビーズ固定ターゲット生体高分子を保存するための第３の溶液をゲルによって互いに区切り、電気泳動により生体高分子を前記第１の溶液中から前記ゲルを経由して前記第２の溶液中へ移動させると共に、ゲル中を泳動中の磁気ビーズ固定ターゲット生体高分子を磁気吸引して前記第３の溶液中に移動させることにより磁気ビーズ固定ターゲット生体高分子の抽出を行うことを特徴とする。
【００１０】
電気泳動と磁気吸引の組合わせにより、磁気ビーズ固定ターゲット生体高分子を容易に抽出することができる。従来のような溶液の移動は一切伴わず、ポンプやバルブ、ミキサーなどを必要としない。
【００１１】
この場合、請求項２のように、ゲルに代えて微小円柱群または多孔質フィルターなどを使用することができる。
【００１２】
請求項３の装置の発明は、生体試料から負の電荷を帯び磁気ビーズに固定されたターゲット生体高分子を抽出する装置であって、
前記生体試料を含む第１の溶液と、移動された生体高分子を保存するための第２の溶液と、抽出された磁気ビーズ固定ターゲット生体高分子を保存するための第３の溶液と、これら３つの溶液を互いに区切るゲルとを保持した容器と、
この容器内に、前記第１の溶液中からゲルおよび第２の溶液の方へ、負の電荷を帯びた生体高分子を電気泳動により移動させるために設けられた正負の電極と、
この正負の電極に正負の電圧をそれぞれ印加する電源と、
電気泳動により前記ゲル中を移動中の磁気ビーズ固定ターゲット生体高分子を磁気吸引して前記第３の溶液中へ移動させるための磁場を発生する磁界発生手段を備え、電気泳動および磁気吸引により磁気ビーズ固定ターゲット生体高分子を第３の溶液中に移動し抽出することができるように構成したことを特徴とする。
【００１３】
このような構成によれば、電気泳動と磁気吸引の組合わせにより、磁気ビーズ固定ターゲット生体高分子を容易に抽出することができる。そして、従来のような溶液の移動は一切伴わず、ポンプやバルブ、ミキサーなどを必要としないため、装置の小型化も可能である。
【００１４】
この場合、請求項４のように、ゲルに代えて微小円柱群または多孔質フィルターなどを使用することができる。
また、請求項５のように、磁界発生手段としては、電磁石または電磁コイルまたは永久磁石を使用することができる。
【００１５】
【発明の実施の形態】
以下図面を用いて本発明を詳しく説明する。図１は本発明に係る磁気ビーズに固定の生体高分子の抽出方法を実施するための装置の一実施例を示す要部構成図である。本発明では、生体試料である生体高分子［例えばDNAやRNA（RNAはDNAからの転写産物、すなわちmRNAまたはrRNAまたはtRNAまたは低分子RNAである）、タンパク質などである］の中から、負の電荷を帯びた既知のターゲット生体高分子を抽出するものである。通常の電気泳動装置により未知の生体高分子を電気泳動により画定・同定するものとは相違する。
【００１６】
なお、本実施例では、生体高分子としてＤＮＡ（より詳しくはＤＮＡ断片）を例にとって説明する。図において、１はガラス板などで平型箱状に形成され、ＤＮＡの電気泳動を行うための密閉状の容器である。この容器１内には、中央部にゲル４が配置されると共に、そのゲル４の三方に接して、生体試料を含む溶液（溶液Ａあるいは第１の溶液ともいう）２と、移動された生体高分子を保存するための溶液（溶液Ｂあるいは第２の溶液ともいう）３と、抽出されたターゲットＤＮＡ５を保存するための溶液（溶液Ｃあるいは第３の溶液ともいう）１０がそれぞれ充填されている。
【００１７】
なお、ターゲットＤＮＡ５は負の電荷を帯び、磁気ビーズ５ａ表面には複数個のターゲットＤＮＡ５が固定されている。なお、ここでは、磁気ビーズに固定されたターゲットＤＮＡを磁気ビーズ固定ターゲットＤＮＡともいう。
【００１８】
また、溶液Ａと溶液Ｂ中にはそれぞれ負の電極（−電極という）６と正の電極（＋電極という）７が配置されており、この２つの電極には電源８から負と正の電圧がそれぞれ印加される。
また、容器１の溶液Ｃ部の外側には磁気ビーズを磁気吸引するための磁場を発生する磁界発生手段１１が配置されている。
【００１９】
このような構成における動作を次に説明する。溶液Ａ中に生体試料を注入しておく。生体試料には、磁気ビーズ固定ターゲットＤＮＡ５と、それ以外の生体高分子が混在している。この生体試料中からターゲットＤＮＡ５を以下のようにして抽出する。
【００２０】
まず、＋電極７と−電極６に電源８からの正負の電圧をそれぞれ印加して電気泳動を行う。負の電荷を帯びているターゲットＤＮＡ５および他の生体高分子は＋電極７側に引き寄せられ、移動する。
一方、同時に磁界発生手段１１により溶液Ｃの方向に磁界をかけると、ゲル４中を泳動中の磁気ビーズ結合ターゲットＤＮＡ５は溶液Ｃ中へ引き寄せられ、分離抽出される。電気泳動中の他の生体高分子は磁気を帯びていないので、磁界に影響されず、溶液Ｂ中へ移動する。
【００２１】
なお、本発明は、上記実施例に限定されることなく、その本質から逸脱しない範囲で更に多くの変更、変形をも含むものである。
例えば、ゲルに代えて、微小な柱（ピラー）のアレイあるいは多孔質で形成されたフィルターを使用してもよい。
【００２２】
また、実施例ではＤＮＡを例にとって説明したが、本発明はＤＮＡに限らず、負の電荷を帯び磁気ビーズに結合された生体高分子を対象に抽出することができる。
また、磁界発生手段として、電磁石または電磁コイルまたは永久磁石を用いることもできる。
【００２３】
【発明の効果】
以上説明したように本発明によれば、μＴＡＳ活用のデバイスで必要としたポンプやバルブ、ミキサーなどを用いることなく、また溶液などの移動を伴うことなく、電気泳動と磁気吸引とによって生体試料から磁気ビーズ固定ターゲット生体高分子を容易に分離し抽出することができる。
【００２４】
なお、μＴＡＳ技術を応用したデバイスは今後色々と実用化されて来る。そのとき、成分の分離や抽出が目的であり、しかも対象が電荷を帯びているような場合には、電気泳動により抽出を行うことが可能であり、機構が複雑となるポンプやバルブを使わなくてもその目的を達成することができる箇所には本発明を適用することができ、その効果は大である。
【００２５】
また、細胞から分子あるいは生体高分子を抽出する抽出装置や前処理装置、あるいは抽出機能およびＤＮＡ増幅機能そして検出反応を一体型で行うようにしたカートリッジなどにおいては、分子あるいは生体高分子の中から磁気ビーズに固定のターゲット分子を分離・抽出する部分に本発明を適用することができる。
【図面の簡単な説明】
【図１】本発明に係る磁気ビーズに固定の生体高分子の抽出方法を実施するための装置の一実施例を示す要部構成図である。
【符号の説明】
１ 容器
２ 溶液Ａ
３ 溶液Ｂ
４ ゲル
５ 磁気ビーズ固定ターゲットＤＮＡ
６、７ 電極
８ 電源
１０ 溶液Ｃ
１１ 磁界発生手段[0001]
BACKGROUND OF THE INVENTION
INDUSTRIAL APPLICABILITY The present invention is used in an extraction device that extracts and separates biopolymers from cells of a biological sample, a pretreatment device thereof, or a cartridge that integrally performs extraction, amplification, detection, etc. of biopolymers from cells. In particular, the present invention relates to a method and apparatus for separating and extracting a target biopolymer from a biopolymer that is a biological sample.
[0002]
[Prior art]
Hereinafter, DNA will be described as an example of a biopolymer. There are roughly two types of methods for extracting and recovering target DNA used for DNA chips from living cells. One is a method using centrifugation, and the other is a method using magnetic beads.
Since the method for centrifugal separation is a large-scale apparatus, the method using magnetic beads is expected to become the mainstream in the future direction where downsizing is desired (see, for example, Non-Patent Document 1 as an application example of magnetic beads). .
[0003]
The method using magnetic beads is, for example, the following method. After immobilizing probe DNA or probe antibody at a certain density on the surface of the magnetic beads, recovering the DNA in the solution by complementary binding of the target DNA and the probe in the solution, and then collecting and washing the magnetic beads with a magnet In this method, DNA is dissociated from the surface of the magnetic beads and recovered.
[0004]
Now, as a device employing such a magnetic bead system, a device about the size of a desktop personal computer has been realized, but no device that has been reduced to a chip size has yet appeared.
However, devices that utilize μTAS (micro / miniaturized total analysis system) for miniaturization have been introduced in various fields (for example, see Non-Patent Document 2 for μTAS).
[0005]
[Non-Patent Document 1]
Supervised by Zenaga Matsunaga, "DNA chip application technology", CMC Co., Ltd., issued July 2000, Chapter 7 DNA chip using magnetic beads Haruko Takeyama, Hideki Nakayama [Non-patent Document 2]
Shoichi Shoko, “Biochemical Microchemical Analysis System-Micromachine Technology-”, page 1, items 1 and 2, Fig. 1, [Search 2003/02/26], Internet <URL: http: // www. jaclap.org/LabCP/p11.html>
[0006]
[Problems to be solved by the invention]
However, in this μTAS device, the pump that is the driving device, the valve that is the control device, and the mixer that is the agitation device have not been realized in practical use. There aren't many that have been made.
[0007]
This is thought to be because in the micro world, fluid dynamic characteristics change greatly depending on fluid viscosity, flow channel shape, etc., and elemental technologies that can solve problems economically and functionally are still in the trial and error stage.
Therefore, the appearance of a system that can separate and extract a target biopolymer fixed to a magnetic bead from other biopolymers without causing fluid movement is desired.
[0008]
An object of the present invention is to solve the above-described problems, and to separate and extract a target biopolymer fixed to a magnetic bead from another biopolymer (hereinafter simply referred to as extraction) without accompanying fluid movement. It is an object of the present invention to provide a biopolymer extraction method and apparatus fixed to magnetic beads that can be miniaturized and can be reduced in size.
[0009]
[Means for Solving the Problems]
In order to achieve such an object, the method invention of claim 1
A method for extracting a target biopolymer that is negatively charged and fixed to a magnetic bead from a biological sample,
A first solution containing the biological sample, a second solution for storing the transferred biopolymer, and a third solution for storing the extracted magnetic bead-fixed target biopolymer by gel. The biopolymers are separated from each other by electrophoresis and transferred from the first solution to the second solution via the gel, and the magnetic bead-fixed target biopolymer being migrated in the gel is magnetically attracted. Then, the extraction of the magnetic bead-fixed target biopolymer is performed by moving it into the third solution.
[0010]
The magnetic bead-fixed target biopolymer can be easily extracted by a combination of electrophoresis and magnetic suction. There is no movement of the solution as in the prior art, and no pump, valve, mixer, etc. are required.
[0011]
In this case, as in claim 2, a group of microcylinders or a porous filter can be used instead of the gel.
[0012]
The invention of the device of claim 3 is an apparatus for extracting a target biopolymer that is negatively charged and fixed to a magnetic bead from a biological sample,
A first solution containing the biological sample; a second solution for storing the transferred biopolymer; a third solution for storing the extracted magnetic bead-fixed target biopolymer; and A container holding a gel that separates the three solutions from each other;
In this container, positive and negative electrodes provided for moving a negatively charged biopolymer by electrophoresis from the first solution to the gel and the second solution;
A power source for applying positive and negative voltages to the positive and negative electrodes,
Magnetic bead fixing target biopolymer moving through the gel by electrophoresis is provided with magnetic field generating means for generating a magnetic field for magnetically attracting and moving the target biopolymer into the third solution. A feature is that the bead-fixed target biopolymer can be moved into the third solution and extracted.
[0013]
According to such a configuration, the magnetic bead-fixed target biopolymer can be easily extracted by a combination of electrophoresis and magnetic attraction. Since the solution does not move at all as in the prior art and does not require a pump, a valve, a mixer, etc., the apparatus can be downsized.
[0014]
In this case, as in claim 4, a group of microcylinders or a porous filter can be used instead of the gel.
As in the fifth aspect, an electromagnet, an electromagnetic coil, or a permanent magnet can be used as the magnetic field generating means.
[0015]
DETAILED DESCRIPTION OF THE INVENTION
Hereinafter, the present invention will be described in detail with reference to the drawings. FIG. 1 is a block diagram showing the main part of an apparatus for carrying out the method for extracting a biopolymer immobilized on a magnetic bead according to the present invention. In the present invention, negatively selected from biological macromolecules [for example, DNA or RNA (RNA is a transcription product from DNA, ie, mRNA, rRNA, tRNA, or small RNA), protein, etc.] Extracts a known target biopolymer having a charge. This is different from that in which an ordinary electrophoretic device defines and identifies an unknown biopolymer by electrophoresis.
[0016]
In this embodiment, DNA (more specifically, a DNA fragment) will be described as an example of a biopolymer. In the figure, reference numeral 1 denotes a hermetic container for electrophoresis of DNA, which is formed in a flat box shape with a glass plate or the like. In this container 1, a gel 4 is disposed at the center, and a solution (also referred to as a solution A or a first solution) 2 containing a biological sample is in contact with three sides of the gel 4 and a living body moved. A solution (also referred to as a solution B or a second solution) 3 for storing the polymer and a solution (also referred to as a solution C or a third solution) 10 for storing the extracted target DNA 5 are filled, respectively. Yes.
[0017]
The target DNA 5 is negatively charged, and a plurality of target DNAs 5 are fixed on the surface of the magnetic beads 5a. Here, the target DNA fixed to the magnetic beads is also referred to as magnetic bead fixed target DNA.
[0018]
Further, a negative electrode (referred to as a negative electrode) 6 and a positive electrode (referred to as a positive electrode) 7 are arranged in the solution A and the solution B, respectively, and negative and positive voltages are supplied to these two electrodes from a power source 8. Are applied respectively.
A magnetic field generating means 11 for generating a magnetic field for magnetically attracting magnetic beads is disposed outside the solution C part of the container 1.
[0019]
The operation in such a configuration will be described next. A biological sample is injected into the solution A. In the biological sample, the magnetic bead fixed target DNA 5 and other biopolymers are mixed. The target DNA 5 is extracted from this biological sample as follows.
[0020]
First, electrophoresis is performed by applying positive and negative voltages from the power source 8 to the + electrode 7 and the − electrode 6, respectively. Negatively charged target DNA 5 and other biopolymers are attracted and moved to the + electrode 7 side.
On the other hand, when a magnetic field is applied in the direction of the solution C by the magnetic field generating means 11 at the same time, the magnetic bead binding target DNA 5 being migrated in the gel 4 is drawn into the solution C and separated and extracted. Since other biopolymers during electrophoresis are not magnetized, they move into the solution B without being affected by the magnetic field.
[0021]
The present invention is not limited to the above-described embodiments, and includes many changes and modifications without departing from the essence thereof.
For example, instead of gel , an array of minute pillars (pillars) or a porous filter may be used.
[0022]
In the embodiments, DNA has been described as an example. However, the present invention is not limited to DNA, and a biopolymer having a negative charge and bound to magnetic beads can be extracted.
In addition, an electromagnet, an electromagnetic coil, or a permanent magnet can be used as the magnetic field generating means.
[0023]
【The invention's effect】
As described above, according to the present invention, from a biological sample by electrophoresis and magnetic suction without using a pump, a valve, a mixer, etc. required for a device using μTAS, and without moving a solution or the like. The magnetic bead-fixed target biopolymer can be easily separated and extracted.
[0024]
Various devices using the μTAS technology will be put into practical use in the future. At that time, the purpose is separation and extraction of components, and if the target is charged, it can be extracted by electrophoresis without using a pump or valve that complicates the mechanism. However, the present invention can be applied to places where the object can be achieved, and the effect is great.
[0025]
In addition, in an extraction device or pretreatment device that extracts molecules or biopolymers from cells, or a cartridge that performs an extraction function, a DNA amplification function, and a detection reaction in an integrated manner, the molecules or biopolymers can be used. The present invention can be applied to a portion where a target molecule fixed to a magnetic bead is separated and extracted.
[Brief description of the drawings]
BRIEF DESCRIPTION OF DRAWINGS FIG. 1 is a block diagram showing the main part of an embodiment of an apparatus for carrying out a method for extracting a biopolymer immobilized on magnetic beads according to the present invention.
[Explanation of symbols]
1 Container 2 Solution A
3 Solution B
4 Gel 5 Magnetic beads fixed target DNA
6, 7 Electrode 8 Power supply 10 Solution C
11 Magnetic field generation means
Claims (5)
前記生体試料を含む第１の溶液と、移動された生体高分子を保存するための第２の溶液と、抽出された磁気ビーズ固定ターゲット生体高分子を保存するための第３の溶液をゲルによって互いに区切り、電気泳動により生体高分子を前記第１の溶液中から前記ゲルを経由して前記第２の溶液中へ移動させると共に、ゲル中を泳動中の磁気ビーズ固定ターゲット生体高分子を磁気吸引して前記第３の溶液中に移動させることによりターゲット生体高分子の抽出を行うことを特徴とする磁気ビーズに固定の生体高分子の抽出方法。A method for extracting a target biopolymer that is negatively charged and fixed to a magnetic bead from a biological sample,
A first solution containing the biological sample, a second solution for storing the transferred biopolymer, and a third solution for storing the extracted magnetic bead-fixed target biopolymer by gel. The biopolymers are separated from each other by electrophoresis and transferred from the first solution to the second solution via the gel, and the magnetic bead-fixed target biopolymer being migrated in the gel is magnetically attracted. And extracting the target biopolymer by moving it into the third solution, and extracting the biopolymer fixed to the magnetic beads.
前記生体試料を含む第１の溶液と、移動された生体高分子を保存するための第２の溶液と、抽出された磁気ビーズ固定ターゲット生体高分子を保存するための第３の溶液と、これら３つの溶液を互いに区切るゲルとを保持した容器と、
この容器内に、前記第１の溶液中からゲルおよび第２の溶液の方へ、負の電荷を帯びた生体高分子を電気泳動により移動させるために設けられた正負の電極と、
この正負の電極に正負の電圧をそれぞれ印加する電源と、
電気泳動により前記ゲル中を移動中の磁気ビーズ固定ターゲット生体高分子を磁気吸引して前記第３の溶液中へ移動させるための磁場を発生する磁界発生手段を備え、電気泳動および磁気吸引により磁気ビーズ固定ターゲット生体高分子を第３の溶液中に移動し抽出することができるように構成したことを特徴とする磁気ビーズに固定の生体高分子の抽出装置。An apparatus for extracting a target biopolymer that is negatively charged and fixed to a magnetic bead from a biological sample,
A first solution containing the biological sample; a second solution for storing the transferred biopolymer; a third solution for storing the extracted magnetic bead-fixed target biopolymer; and A container holding a gel that separates the three solutions from each other;
In this container, positive and negative electrodes provided for moving a negatively charged biopolymer by electrophoresis from the first solution to the gel and the second solution;
A power source for applying positive and negative voltages to the positive and negative electrodes,
Magnetic bead fixing target biopolymer moving through the gel by electrophoresis is provided with magnetic field generating means for generating a magnetic field for magnetically attracting and moving the target biopolymer into the third solution. An apparatus for extracting a biopolymer fixed to a magnetic bead, wherein the bead-fixed target biopolymer can be moved and extracted into a third solution.
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| JP2003103714A JP4039297B2 (en) | 2003-04-08 | 2003-04-08 | Extraction method and extraction device for biopolymer fixed to magnetic beads |
| US10/801,544 US20040203126A1 (en) | 2003-04-08 | 2004-03-17 | Method and apparatus for separating and purifying biopolymers |
| US11/507,527 US20060286596A1 (en) | 2003-04-08 | 2006-08-22 | Method and apparatus for separating and purifying biopolymers |
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