JP2018128337A - 生体関連分子固定化用担体 - Google Patents
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Abstract
【解決手段】樹脂基板と、該樹脂基板上に形成されたアミノ基含有化合物層と、該アミノ基含有化合物層上に形成された多価カルボン酸層を有する生体関連分子固定化用担体であって、前記多価カルボン酸層のカルボキシル基は活性エステル化されており、前記樹脂基板は無機顔料を含み、かつ、前記樹脂基板の中心線表面平均粗さ Raは60nm以下であることを特徴とする、生体関連分子固定化用担体。
【選択図】図1
Description
以下、実施例に基づいて本願の態様をさらに詳細に説明する。
カーボンブラックを0.5%含有するポリカーボネートを用い、表面粗さRaが異なる基板を射出成形により作製した。表面処理前の素板を、DNAチップ検出機GENOGATE Reader(東洋製罐グループホールディングス社製)を用いて撮影した。撮影条件として、乾燥状態の素板に640nmのレザー光を照射し、16bitモノクロカメラ(輝度範囲:0(黒)〜65536(白))、露光時間15[s]にて撮影し、その際の各素板の3点の画像輝度の平均値をバックグランウド値(BG値)として測定した。結果を表1に示す。
カーボンブラックを0.5%含有するポリカーボネートを用い、表面粗度Raの異なる樹脂基板を作製した。上記1と同様の方法により素板のBG値を測定した。
その後、樹脂基板を5重量%の3−アミノプロピルトリエトキシシラン水溶液に30分間浸漬してアミノ基を導入した。このアミノ基が導入された基板を1重量%ポリアクリル酸水溶液に10分間浸漬した後、純水で洗浄し、0.1Mリン酸緩衝液(pH6.8)に100mMの1−エチル−3−(3−ジメチルアミノプロピル)カルボジイミド塩酸塩と50mMのN−ヒドロキシスクシンイミドを溶解した活性化液中に10分間浸漬することによってカルボキシル基を活性化した担体を作製した。
上記の担体に対し、マイクロアレイ作製装置を用いて10mMに調製したDNAプローブ溶液を点着した。DNAプローブが点着された基板を80℃のオーブンで1時間加熱した後、2×SSC/0.2%SDSで室温にて10分間および60℃にて10分間洗浄してDNA固定化基板を作製した。
ハイブリダイゼーション用バッファー40mlとプローブと相補なDNAを0.125 nMにて混合し、作製したDNA固定化担体の表面と接触させ、45℃で60分間反応させた。反応後、基板を0.5×SSC/0.2%SDS溶液で、次いで0.5×SSC溶液で2回、それぞれ50往復ずつ揺動洗浄し、カバーガラスをのせてGENOGATE Reader(東洋製罐グループホールディングス社製)で蛍光検出画像を得た。検出画像上の各スポットから得られる蛍光強度値およびバックグラウンド値からS/N比((蛍光強度−BG値)/BG値)を算出した。DNAチップの測定は、各実施例及び比較例について4つの16スポットを有する担体を4枚作製し、計64スポットについて測定した平均値を求めた。担体のスポット後の画像を図2に、また、スポット特性及び検出時の測定結果を表2にそれぞれ示す。
上記の結果から、表面粗さを本願所定の範囲とすることにより、スポット形状が良好であり、かつ、BG値の低い担体を得られることが分かる。
Claims (8)
- 樹脂基板と、該樹脂基板上に形成されたアミノ基含有化合物層と、該アミノ基含有化合物層上に形成された多価カルボン酸層を有する生体関連分子固定化用担体であって、前記多価カルボン酸層のカルボキシル基は活性エステル化されており、前記樹脂基板は無機顔料を含み、かつ、前記樹脂基板の中心線表面平均粗さRaは60nm以下であることを特徴とする、生体関連分子固定化用担体。
- 前記樹脂基板の中心線表面平均粗さRaが10nm以下であることを特徴とする、請求項1に記載の生体関連分子固定化用担体。
- 前記無機顔料が黒色顔料である、請求項1又は2に記載の生体関連分子固定化用担体。
- 前記黒色顔料が、カーボンブラック、カーボンナノチューブ,アニリンブラック、チタンブラック、アセチレンブラック、ヘマタイト、ペリレンブラック及びこれらの混合物からなる群から選択されることを特徴とする、請求項3に記載の生体関連分子固定化用担体。
- 前記樹脂基板の中の無機顔料の含有量が、樹脂100重量部に対して0.1〜5重量部であることを特徴とする、請求項1〜4のいずれか1項に記載の生体関連分子固定化用担体。
- 前記樹脂基板の樹脂がポリカーボネート又はポリプロピレンであることを特徴とする、請求項1〜5のいずれか1項に記載の生体関連分子固定化用担体。
- 無機顔料を含み、中心線表面平均粗さRaが60nm以下である樹脂基板を提供する工程、前記樹脂基板の上にアミノアルキルシラン層を形成する工程、該アミノアルキルシラン層の上に多価カルボン酸層を形成する工程、及び、該多価カルボン酸層のカルボキシル基を活性エステル化する工程を含む、生体関連分子固定化用担体の製造方法。
- 前記樹脂基板を、射出成形又は押出し成形により成形することを特徴とする、請求項7に記載の製造方法。
Priority Applications (4)
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JP2017021269A JP6789510B2 (ja) | 2017-02-08 | 2017-02-08 | 生体関連分子固定化用担体 |
PCT/JP2018/004453 WO2018147383A1 (ja) | 2017-02-08 | 2018-02-08 | 生体関連分子固定化用担体 |
EP18751798.2A EP3581930A4 (en) | 2017-02-08 | 2018-02-08 | CARRIER FOR IMMOBILIZATION OF BIOLOGICAL MOLECULES |
US16/483,449 US20210132047A1 (en) | 2017-02-08 | 2018-02-08 | Carrier for bio-related molecule immobilization |
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JP2017021269A JP6789510B2 (ja) | 2017-02-08 | 2017-02-08 | 生体関連分子固定化用担体 |
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JP2018128337A true JP2018128337A (ja) | 2018-08-16 |
JP6789510B2 JP6789510B2 (ja) | 2020-11-25 |
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US (1) | US20210132047A1 (ja) |
EP (1) | EP3581930A4 (ja) |
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Citations (5)
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JP2002211954A (ja) * | 2000-11-07 | 2002-07-31 | Toyo Kohan Co Ltd | 粗面化スライドグラス及びそれを用いて生体物質を解析する方法 |
JP2003139705A (ja) * | 2001-10-30 | 2003-05-14 | Sumitomo Bakelite Co Ltd | マイクロチップ用基板 |
JP2005043312A (ja) * | 2003-07-25 | 2005-02-17 | Toyo Kohan Co Ltd | 真空パック固体支持体アレイ及びその製造方法 |
JP2008249416A (ja) * | 2007-03-29 | 2008-10-16 | Toyo Kohan Co Ltd | マイクロアレイ用基板および該基板を用いたマイクロアレイ |
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DE10233312A1 (de) * | 2002-07-22 | 2004-03-04 | Westfaliasurge Gmbh | Verfahren und Vorrichtung zur Bestimmung eines Volumenstroms während eines Melkvorgangs einer fließenden Milch |
US20050176003A1 (en) * | 2001-11-27 | 2005-08-11 | Sumitomo Bakelite Co., Ltd. | Plastic substrate for microchips |
JP2004097173A (ja) * | 2002-07-17 | 2004-04-02 | Toyo Kohan Co Ltd | 静電層を有する固体支持体及びその用途 |
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JP4689475B2 (ja) | 2006-01-11 | 2011-05-25 | ニプロ株式会社 | 核酸固定用成形体および核酸固定化方法 |
JP5459990B2 (ja) * | 2008-06-30 | 2014-04-02 | 株式会社ハイペップ研究所 | バイオチップ用基板及びその製造方法 |
US9458315B2 (en) * | 2009-08-25 | 2016-10-04 | Sabic Global Technologies B.V. | Flame retardant thermoplastic polycarbonate compositions and films made therefrom |
CN102892848A (zh) * | 2010-05-17 | 2013-01-23 | 日东电工株式会社 | 底涂剂组合物及粘合片 |
JP6111941B2 (ja) * | 2013-09-06 | 2017-04-12 | 日本軽金属株式会社 | バイオチップ用基板 |
CN105793055B (zh) * | 2013-11-28 | 2019-06-07 | 三菱化学株式会社 | 透明激光打标片材、层叠体和激光打标方法 |
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JP2002211954A (ja) * | 2000-11-07 | 2002-07-31 | Toyo Kohan Co Ltd | 粗面化スライドグラス及びそれを用いて生体物質を解析する方法 |
JP2003139705A (ja) * | 2001-10-30 | 2003-05-14 | Sumitomo Bakelite Co Ltd | マイクロチップ用基板 |
JP2005043312A (ja) * | 2003-07-25 | 2005-02-17 | Toyo Kohan Co Ltd | 真空パック固体支持体アレイ及びその製造方法 |
JP2008249416A (ja) * | 2007-03-29 | 2008-10-16 | Toyo Kohan Co Ltd | マイクロアレイ用基板および該基板を用いたマイクロアレイ |
WO2011030823A1 (ja) * | 2009-09-10 | 2011-03-17 | 東洋鋼鈑株式会社 | 核酸を保持するための担体 |
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Publication number | Publication date |
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EP3581930A1 (en) | 2019-12-18 |
JP6789510B2 (ja) | 2020-11-25 |
WO2018147383A1 (ja) | 2018-08-16 |
US20210132047A1 (en) | 2021-05-06 |
EP3581930A4 (en) | 2020-12-16 |
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