JP2005274141A - 膜たんぱく質固定化基板および固定化方法 - Google Patents
膜たんぱく質固定化基板および固定化方法 Download PDFInfo
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Abstract
【解決手段】 基板の表面に、変性タンパク質の単分子膜を付着させる(S101)。変性タンパク質として、膜タンパク質が変性したタンパク質を用いる。膜の表面に、被固定化物質を接着させるために、特定物質に対して特異的相互作用する親和性物質を固定化する(S102)。そして、親和性物質を固定化した膜の表面に、特定物質を固定化する(S103)。基板表面に固定化させたい被固定化物質に親和性物質を結合させておき、基板の表面に固定化された固定化物質に特異的に吸着させる(S104)。
【選択図】 図1
Description
図7は、前述のように、被固定化物質がタンパク質である場合を示す図である。ビオチン283−アビジン285−ビオチン284のサンドイッチ構造を介して変性タンパク質単分子膜352にタンパク質274を固定化することにより、タンパク質274の変性を抑制し、安定的に固定化することができる。このような固定化基板は、たとえば表面プラズモン測定用のセンサチップ、AFM測定用の基板、その他各種バイオセンサに好適に利用できる。
第1の実施の形態では、変性タンパク質単分子膜352と被固定化物質との接続に、ビオチン283とアビジン285を用いたが、これらに代えて、Ni(II)とNTAおよびヒスチジンペプチドの組み合わせを用いてもよい。
Claims (7)
- 基板と、
前記基板上に設けられた膜タンパク質またはその変性タンパク質を含む膜と、
前記膜の表面に設けられ、特定物質に選択的に吸着する親和性物質と、
を有することを特徴とする固定化基板。 - 請求項1に記載の固定化基板において、
前記親和性物質が第一の親和性物質および第二の親和性物質を含み、
前記第一の親和性物質は前記膜の表面に結合し、
前記特定物質は前記第一の親和性物質および前記第二の親和性物質に選択的に吸着し、
前記第二の親和性物質に結合している被固定化物質を有することを特徴とする固定化基板。 - 請求項1または2に記載の固定化基板において、前記膜が前記タンパク質またはその変性タンパク質の単分子膜または前記単分子膜の積層膜であることを特徴とする固定化基板。
- 請求項1乃至3いずれかに記載の固定化基板において、前記膜タンパク質がバクテリオロドプシンであることを特徴とする固定化基板。
- 表面に変性膜タンパク質を含む膜を有する基板を準備するステップと、
特定物質に選択的に吸着する第一の親和性物質を前記膜の表面に固定化するステップと、
前記特定物質に選択的に吸着する第二の親和性物質が結合した被固定化物質を含む液体を準備するステップと、
前記第一の親和性物質または前記第二の親和性物質に前記特定物質を吸着させるステップと、
前記被固定化物質を含む液体を、前記膜の表面に展開し、前記膜の表面に前記被固定化物質を固定化するステップと、
を含むことを特徴とする固定化方法。 - 請求項5に記載の固定化方法において、基板を準備する前記ステップは、膜タンパク質の分散液または溶液を液体表面に展開し、前記膜タンパク質を変性させて、前記変性膜タンパク質を含む単分子膜を形成するステップと、
前記単分子膜を前記基板の表面に付着させるステップと、
を含むことを特徴とする固定化方法。 - 請求項6に記載の固定化方法において、前記膜タンパク質がバクテリオロドプシンであることを特徴とする固定化方法。
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JP2004083321A JP4484553B2 (ja) | 2004-03-22 | 2004-03-22 | 膜たんぱく質固定化基板および固定化方法 |
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JP2005274141A true JP2005274141A (ja) | 2005-10-06 |
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Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2009008556A1 (ja) * | 2007-07-11 | 2009-01-15 | National Institute For Materials Science | 柔らかで自立性があるタンパク質ナノ薄膜、その製造法及び応用 |
JP2011226920A (ja) * | 2010-04-20 | 2011-11-10 | National Institute Of Advanced Industrial & Technology | ポリマー脂質二分子膜を用いた機能性基板 |
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JPH05223815A (ja) * | 1992-02-07 | 1993-09-03 | B M L:Kk | 免疫グロブリンgの微量測定法 |
JPH10182693A (ja) * | 1996-11-05 | 1998-07-07 | Seibutsu Bunshi Kogaku Kenkyusho:Kk | 分子の配向制御方法 |
WO2002057780A1 (en) * | 2001-01-18 | 2002-07-25 | Newcastle University Ventures Ltd. | Biosensor with covalently attached membrane-spanning proteins |
WO2002072873A1 (en) * | 2001-03-14 | 2002-09-19 | Biacore Ab | Method of preparing supported lipid film membranes and use thereof |
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Patent Citations (5)
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JPH01248570A (ja) * | 1988-03-30 | 1989-10-04 | Agency Of Ind Science & Technol | アビジン―ビオチン系を用いた光電変換装置とその製造方法 |
JPH05223815A (ja) * | 1992-02-07 | 1993-09-03 | B M L:Kk | 免疫グロブリンgの微量測定法 |
JPH10182693A (ja) * | 1996-11-05 | 1998-07-07 | Seibutsu Bunshi Kogaku Kenkyusho:Kk | 分子の配向制御方法 |
WO2002057780A1 (en) * | 2001-01-18 | 2002-07-25 | Newcastle University Ventures Ltd. | Biosensor with covalently attached membrane-spanning proteins |
WO2002072873A1 (en) * | 2001-03-14 | 2002-09-19 | Biacore Ab | Method of preparing supported lipid film membranes and use thereof |
Cited By (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2009008556A1 (ja) * | 2007-07-11 | 2009-01-15 | National Institute For Materials Science | 柔らかで自立性があるタンパク質ナノ薄膜、その製造法及び応用 |
JP2009131725A (ja) * | 2007-07-11 | 2009-06-18 | National Institute For Materials Science | 柔らかで自立性があるタンパク質ナノ薄膜、その製造法及び応用 |
EP2179779A3 (en) * | 2007-07-11 | 2011-11-02 | National Institute for Materials Science | A flexible free-standing ultrathin or thin protein membrane, its fabrication method and application |
CN101918119B (zh) * | 2007-07-11 | 2013-07-03 | 独立行政法人物质·材料研究机构 | 柔软的自立性的蛋白质纳米薄膜、其制造方法和应用 |
CN101862609B (zh) * | 2007-07-11 | 2013-07-03 | 独立行政法人物质·材料研究机构 | 柔软的自立性的蛋白质纳米薄膜、其制造方法和应用 |
US8741152B2 (en) | 2007-07-11 | 2014-06-03 | National Institute For Materials Science | Flexible free-standing ultrathin or thin protein membrane, its fabrication method and application |
US8828239B2 (en) | 2007-07-11 | 2014-09-09 | National Institute For Materials Science | Flexible free-standing ultrathin or thin protein membrane, its fabrication method and application |
JP2011226920A (ja) * | 2010-04-20 | 2011-11-10 | National Institute Of Advanced Industrial & Technology | ポリマー脂質二分子膜を用いた機能性基板 |
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