JP2005233955A - 生体分子を固体基板上に非共有的に固定化する方法及びそれによって製造されるマイクロアレイ - Google Patents
生体分子を固体基板上に非共有的に固定化する方法及びそれによって製造されるマイクロアレイ Download PDFInfo
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Abstract
【解決手段】 水素供与能を有する第1官能基を表面に有する固体基板上で、水素受容能を有する化合物と、水素供与能を有する第2官能基で官能基化された生体分子との混合物を反応させて、前記生体分子を非共有結合によって前記基板上に固定化する段階を含むことを特徴とする。
【選択図】 なし
Description
本実施例では、GAPSでコーティングされたガラス基板上に、PEGと、アミノ基で5’末端が官能基化されたDNAとを利用して、DNAが前記ガラス基板上にスポットで配列されたDNAマイクロアレイを製造した。
本実施例では、実施例1に製造されたプローブポリヌクレオチドマイクロアレイを利用して、プローブポリヌクレオチドと標的核酸とのハイブリダイゼーションによりその結果を検出することによって、本発明の方法によって製作されたマイクロアレイの品質を評価した。
本実施例では、他の条件を実施例1及び2の条件と同一にし、固定化されたプローブヌクレオチドの配列を異ならせてプローブポリヌクレオチドアレイを製作した後、ハイブリダイゼーションを行った。本実施例で使用したプローブポリヌクレオチドの配列は配列番号25と同じく、標的ポリヌクレオチドは、配列番号15に完全相補的なヌクレオチド配列を持つものを使用した。
本実施例では、固体基板としてシリコンウェーハ(LG SIltron社製のホウ素ドーピングされたP型シリコンウェーハ)にGAPSコーティングしたもの使用したことを除いて、実施例1と同じ条件で製作したマイクロアレイで標的核酸をハイブリダイズした後、蛍光強度を測定した。使用したプローブ及び標的ポリヌクレオチドは実施例3と同一であった。
本実施例では、固定化に使われるプローブポリヌクレオチドの種類とその濃度とを変更したことを除いて、実施例1及び2に記載するようにマイクロアレイを製作し、標的核酸をハイブリダイズした後、蛍光強度を測定した。プローブポリヌクレオチドは、20μM及び100μMの濃度で固定化反応に使用した。本実施例に使われたプローブポリヌクレオチドの配列は配列番号26と同じである。
Claims (10)
- 水素供与能を有する第1官能基を表面に有する固体基板上で、水素受容能を有する化合物と、水素供与能を有する第2官能基で官能基化された生体分子との混合物を反応させて、前記生体分子を非共有結合によって前記基板上に固定化する段階を含む、生体分子を固体基板上に固定化する方法。
- 前記水素供与能を有する第1官能基と第2官能基とは、同一であるか、または異なることを特徴とする請求項1に記載の方法。
- 前記水素供与能を有する第1官能基と第2官能基とは、アミノ基、チオール基及びヒドロキシル基よりなる群から選択されることを特徴とする請求項1または2に記載の方法。
- 前記水素受容能を有する化合物は、ポリエチレングリコール(PEG)またはその誘導体、およびポリエチレンイミンまたはその誘導体であることを特徴とする請求項1〜3のいずれかに記載の方法。
- 前記生体分子は、核酸、蛋白質、多糖類及びその組合わせよりなる分子からなる群から選択されるものであることを特徴とする請求項1〜4のいずれかに記載の方法。
- 前記固体基板は、γ−アミノプロピルトリエトキシシラン(GAPS)でコーティングされており、前記水素受容能を有する化合物はポリエチレングリコールであり、前記生体分子は5’末端がアミノアルキルで官能基化されたDNAであることを特徴とする、請求項1に記載の生体分子を固体基板上に固定化する方法。
- 前記ポリエチレングリコールの分子量は、200ないし1,000,000Daであることを特徴とする請求項6に記載の方法。
- 前記ポリエチレングリコールの濃度は、0.4mMないし8.0mMであることを特徴とする請求項6に記載の方法。
- 前記DNAの濃度は20μMないし100μMであることを特徴とする請求項6〜8のいずれかに記載の方法。
- 請求項1ないし9のいずれか1項に記載の方法によって製造されるマイクロアレイ。
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
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KR1020040009942A KR100580644B1 (ko) | 2004-02-16 | 2004-02-16 | 생물분자를 고체 기판상에 비공유적으로 고정화 하는 방법및 그에 의하여 제조되는 마이크로어레이 |
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JP2005233955A true JP2005233955A (ja) | 2005-09-02 |
JP4137898B2 JP4137898B2 (ja) | 2008-08-20 |
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JP2005038463A Expired - Fee Related JP4137898B2 (ja) | 2004-02-16 | 2005-02-15 | 生体分子を固体基板上に非共有的に固定化する方法及びそれによって製造されるマイクロアレイ |
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Country | Link |
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US (2) | US7611889B2 (ja) |
EP (1) | EP1612277B1 (ja) |
JP (1) | JP4137898B2 (ja) |
KR (1) | KR100580644B1 (ja) |
CN (1) | CN100408697C (ja) |
DE (1) | DE602005012183D1 (ja) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
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WO2008093647A1 (ja) * | 2007-01-31 | 2008-08-07 | Tohoku University | マイクロアレイとその製造方法並びに有機分子と作用物質との相互作用の検出方法 |
JP2018501803A (ja) * | 2014-12-31 | 2018-01-25 | コーニンクレッカ フィリップス エヌ ヴェKoninklijke Philips N.V. | 物理吸着を介したポリマー基板上への修飾されたオリゴヌクレオチドの固定化 |
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KR100738083B1 (ko) * | 2005-12-20 | 2007-07-12 | 삼성전자주식회사 | 마이크로어레이용 기판 및 그의 제조방법 |
KR100804713B1 (ko) | 2006-10-30 | 2008-02-18 | 포항공과대학교 산학협력단 | 당류의 직접적인 표면 고정화 방법 |
KR100836206B1 (ko) * | 2007-03-20 | 2008-06-09 | 연세대학교 산학협력단 | 패턴형성용 폴리에틸렌글리콜 하이드로젤과 이의 제조방법 |
ES2625874T3 (es) * | 2008-08-22 | 2017-07-20 | Denka Seiken Co., Ltd. | Dispositivo de prueba para ensayo de membrana que comprende una sección de presentación visual de referencia |
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US5252743A (en) * | 1989-11-13 | 1993-10-12 | Affymax Technologies N.V. | Spatially-addressable immobilization of anti-ligands on surfaces |
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- 2005-02-16 DE DE602005012183T patent/DE602005012183D1/de active Active
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Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2008093647A1 (ja) * | 2007-01-31 | 2008-08-07 | Tohoku University | マイクロアレイとその製造方法並びに有機分子と作用物質との相互作用の検出方法 |
JP2018501803A (ja) * | 2014-12-31 | 2018-01-25 | コーニンクレッカ フィリップス エヌ ヴェKoninklijke Philips N.V. | 物理吸着を介したポリマー基板上への修飾されたオリゴヌクレオチドの固定化 |
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US7687259B2 (en) | 2010-03-30 |
JP4137898B2 (ja) | 2008-08-20 |
US7611889B2 (en) | 2009-11-03 |
CN1821424A (zh) | 2006-08-23 |
KR100580644B1 (ko) | 2006-05-16 |
CN100408697C (zh) | 2008-08-06 |
EP1612277A1 (en) | 2006-01-04 |
KR20050081609A (ko) | 2005-08-19 |
US20080280782A1 (en) | 2008-11-13 |
EP1612277B1 (en) | 2009-01-07 |
US20050181407A1 (en) | 2005-08-18 |
DE602005012183D1 (de) | 2009-02-26 |
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