JP2004309494A - 病原性微生物の分類学的同定及び病原性微生物の毒性タンパク質 - Google Patents
病原性微生物の分類学的同定及び病原性微生物の毒性タンパク質 Download PDFInfo
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Abstract
【解決手段】リガンドは基材の表面から幾分離れた位置に共有結合的にテザーされる。微生物の結合を目的としたリガンドの場合のテザーは少なくとも15Å、タンパク質の結合を目的としたリガンドの場合のテザーは少なくとも6Åであることが必要である。本リガンドとしては、毒性タンパク質、外膜タンパク質及びコンジュゲート脂質に特異的な、ヘム化合物、シデロフォア、多糖類及びペプチドを含む。分析に付される溶液中の非結合成分は結合画分から分離され、結合は、顕微鏡法、蛍光法、エピ蛍光法、ルミネッセンス法、燐光法、放射能法、あるいは吸光度法による検出が可能である。基材表面に多くのリガンドをアレイ状にパターン化することによって、アレイに対する試料の結合パターンを分析すれば、微生物の分類学的同定が可能である。
【選択図】図1
Description
GADRSYLSFIHLYPELAGAGGGC
をテザーしておけば、その末端システイン基によって、特にフリーのStaphylococcus aureus トキシックショック・トキシン−1を捕捉できる。また、次のペプチド、
GHHKHHHGGGC
も、これをテザーしておくことにより、その末端システイン基によって、Staphylococcus aureusの表面に露出しているプロテインAを、よって結局この生物自体を、特異的に捕捉できる。Staphylococcus aureus トキシックショック症候群トキシン−1結合ペプチドについては、A.Satoらによる「ファージディスプレイライブラリーからの、トキシックショック症候群トキシン−1に結合しその主要組織親和性複合体(MHC)クラスII分子との結合を阻害するペプチドの同定(Identification from a Phage Display Library of Peptides that Bind to Toxic Shock Syndrome Toxin−1 and that Inhibit Its Binding to Major Histocompatibility Complex (MHC) Class II Molecules)」(Biochemistry 35、10441−10447(1996))に記載がある。
セクション位置 リガンド(3×3の配列)
A1 アシアロGM1
A2 ヘミン
A3 ピオチェリン
B1 Ga1NAcβGa1
B2 アルカリジン
B3 フィブロネクチン(ペプチド断片)
C1 抗S. aureus プロテイン A ペプチド
C2 スタフィロフェリン
C3 フェロキサミン B
Claims (13)
- 生物学的分析対象物の分類学的同定のための方法であって、
(a)分析対象物を含有する溶液を、マーカーに結合させた所望の分析対象物に対して特異的なリガンドに曝露するステップと、
(b)結合した分析対象物を、過剰なマーカー結合リガンドから分離するステップと、
(c)結合したマーカーの検出によりリガンド結合に関して分析対象物を調べるステップとを含む方法。 - 請求項1記載の方法であって、生物学的分析対象物が、
(a)細菌と、
(b)ウイルスと、
(c)タンパク質トキシンと、
(d)リケッチアと、
(e)原生動物と、
(f)菌類と、
(g)細胞質タンパク質とから成る群から選択される方法。 - 請求項1記載の方法であって、結合した分析対象物の過剰な結合リガンドからの分離がクロマトグラフィーによって達成される方法。
- 請求項1記載の方法であって、前記リガンドが磁気粒子に結合しており、結合した分析対象物の、分析対象物溶液中の非結合成分からの分離が、磁気分離によって達成され、ここにおいて前記リガンドは、前記磁気粒子に、少なくとも15Å離れた位置にテザーされて微生物を捕捉する方法。
- 請求項1記載の方法であって、前記リガンドがヘム化合物である方法。
- 請求項1記載の方法であって、前記リガンドがシデロフォアである方法。
- 請求項1記載の方法であって、前記リガンドが多糖類である方法。
- 請求項1記載の方法であって、前記リガンドが外膜タンパク質に特異的なペプチドである方法。
- 請求項1記載の方法であって、前記リガンドがコンジュゲート脂質に特異的なペプチドである方法。
- 請求項1記載の方法であって、マーカーが蛍光性で検出が蛍光による方法。
- 請求項1記載の方法であって、マーカーが発光性で検出がルミネッセンスによる方法。
- 請求項1記載の方法であって、マーカーが放射性で検出が放射能による方法。
- 請求項1記載の方法であって、マーカーが燐光性で検出が燐光による方法。
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US09/999,159 US6780602B2 (en) | 2001-11-01 | 2001-11-01 | Taxonomic identification of pathogenic microorganisms and their toxic proteins |
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HK1054054A1 (zh) | 2003-11-14 |
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US6780602B2 (en) | 2004-08-24 |
SG130937A1 (en) | 2007-04-26 |
JP2004317520A (ja) | 2004-11-11 |
SG152039A1 (en) | 2009-05-29 |
EP1308520A3 (en) | 2003-11-12 |
SG143054A1 (en) | 2008-06-27 |
US20040096910A1 (en) | 2004-05-20 |
JP2004333503A (ja) | 2004-11-25 |
US20050026140A1 (en) | 2005-02-03 |
CN1417347A (zh) | 2003-05-14 |
US20030124532A1 (en) | 2003-07-03 |
SG152902A1 (en) | 2009-06-29 |
US20050026141A1 (en) | 2005-02-03 |
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