JP2010533871A - 標的分子を検出するためのアレイ、基板、装置、方法、及び、システム - Google Patents
標的分子を検出するためのアレイ、基板、装置、方法、及び、システム Download PDFInfo
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Abstract
Description
(a)検出することができる生体分子の、アッセイの感度又は最小限の濃度、
(b)生体分子を検出することができる濃度範囲、
(c)同時に検出することができる異なる生体分子の数、
(d)測定間での可変性、
(e)同時に検出することができる(例えばmRNA、蛋白質等の)異なる種類の生体分子の数、
(f)測定に必要とされる最小限の試料サイズ、及び
(g)測定を行うことができる速度、
を含む。
図13のパネルAに概略的に例示された手順に従い、バーコードチップを作製した。
DEALバーコードアレイのマイクロチャネルにより誘導されたフローパターン形成の間、ポリ−L−リシン(ポリアミン)による処理によってガラスの表面を修飾し、DNA吸着のための3次元マトリックスを生じて、DNA荷重の量を著しく高めた。
DEAL技術と統合されたバーコードチップは多重化された蛋白質測定に対する高密度アレイを与えるということが、本出願人により行われた一連の実験によって示された。さらに、DEALバーコードチップは、従来のピンスポットマイクロアレイと比較して、感度の著しい改善も実証する。
DNAの検出に対するバイオアッセイにおいてバーコードアレイを使用した。特に、ポリヌクレオチド(DNA)を基板上でパターン形成し、試料内の相補的なポリヌクレオチドを検出するために使用した。パターン形成されたDNAオリゴマーがその相補鎖に結合するために高い親和性を示すことが、図16に例示されている結果によって示されている。
本明細書において開示されるように組み立てたバーコードアレイを、本出願人によって開発された実験法に従い、蛋白質検出のために使用した。
実施例3のバーコードアレイ、及び、ピンスポット技術を使用して印刷された従来のマイクロアレイにおいて比較実験を行った。図15パネルdに例示された結果は、どれ程明瞭に、従来のマイクロアレイがDEALバーコードチップよりも1〜20倍低い感度を得ただけかを示している。
図19に例示されているように、DEAL技術と統合させたバーコードアレイを使用して多数の蛋白質を検出した。特に、図19は、5つの異なる蛋白質の検出に対するDEALバーコードイムノアッセイの使用を示している。前記蛋白質は、従来のスポットマイクロアレイを使用した1つの蛋白質の検出に必要とされる領域よりも小さい領域内で検出される。
図21に例示されているように、DEAL技術と統合させたバイオバーコードを使用してバイオマーカーを検出した。特に、図21は、DEAL技術と共に利用された場合に広げられたバーコードアレイのダイナミックレンジを例示している。データは、40倍を超える広大なダイナミックレンジをカバーすることができるDEALバーコードイムノアッセイを使用した、ヒト血清内の、妊娠検査のマーカーであるhCGの測定を示している。
National Cancer Institute(NCI)により提供された、一連の標準的なヒト絨毛性ゴナドトロピン(hCG)を加えたヒト血清試料に対して検査を行った。hCGは、妊娠検査に対して広く使用されており、妊娠性トロホブラスト腫瘍、並びに、卵巣及び精巣の胚細胞癌に対するバイオマーカーとしても役に立っている。
図23に例示されているように、バーコードアレイを使用して生物学的プロファイルを検出した。特に、図23は、ヒト血清蛋白質プロファイリングに対する統合されたマイクロ流体DEALバーコード装置の使用を示している。12人の癌患者由来の血清試料を、そのようなプロトタイプの臨床検査プラットフォーム(prototype clinic test platform)において測定した。
臨床血液試料に対するバーコードアレイの有用性及び再現性をさらに評価するために、24人の癌患者からの少量の血清由来の12個の蛋白質のパネルをDEALバーコードマイクロ流体装置において測定した。このパネル中の蛋白質は、前立腺特異抗原(PSA)、並びに、種々の白血球によって分泌された11個の蛋白質を含んだ。各血清試料に対して、各バーコードを何度も測定した。
実施例10に例示された実験を通じて測定した血液バーコードは、各患者に特有であった。
血漿蛋白質アッセイに対する改善された感度を確証するよう、バーコードアレイの使用を検査した。
指輪等の小物に対する磁気IDバーコードを製造する方法の概略図が図31に示されている。
政府基金に関する記述
米国政府は、National Institutes of Healthにより与えられた基金No.CA119347のもとに行われた本開示についてある種の権利を有している。
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Claims (33)
- 基板に付着される少なくとも1つの捕獲剤又はその構成成分を含む、試料内の少なくとも1つの標的を検出するためのアレイであって、前記少なくとも1つの捕獲剤が、前記少なくとも1つの標的に特異的に結合して捕獲剤標的結合複合体を形成する能力を持ち、バーコードパターンを形成する実質的に平行な線に沿って捕獲剤標的結合複合体が検出可能であるように前記少なくとも1つの捕獲剤又はその構成成分が当該アレイ上に配置される、アレイ。
- 前記少なくとも1つの標的は複数の標的であり、前記少なくとも1つの捕獲剤又はその構成成分は複数の捕獲剤又はその構成成分であり、前記複数の捕獲剤の各捕獲剤は、互いから結合的に区別可能且つ位置的に区別可能であり、前記複数の捕獲剤の各捕獲剤は、前記複数の標的の各標的に特異的に結合して捕獲剤標的結合複合体を形成する能力を持つ、請求項1に記載のアレイ。
- 前記複数の標的が複数のバイオマーカーを含む、請求項2に記載のアレイ。
- 前記バーコードパターンが生物学的プロファイルと付随する、請求項3に記載のアレイ。
- 前記生物学的プロファイルが、疾患と付随する所定の生物学的プロファイルとの比較により診断指標を提供する、請求項4に記載のアレイ。
- 前記実質的に平行な線がマイクロ流体チャネル又はその一部によって形成され、前記マイクロ流体チャネルが当該アレイのマイクロ流体チャネルである、請求項1乃至5のいずれか一項に記載のアレイ。
- 前記複数の捕獲剤又はその構成成分が、
当該アレイに付着される複数のアレイポリヌクレオチドであり、該アレイに付着される複数のアレイポリヌクレオチドの各ポリヌクレオチドが配列特異的で且つ互いから位置的に区別可能である、アレイポリヌクレオチドを含む、請求項2乃至5のいずれか一項に記載のアレイ。 - 前記複数の捕獲剤又はその構成成分が、
複数のポリヌクレオチドによりコード化される蛋白質であり、各ポリヌクレオチドによりコード化される蛋白質が、蛋白質、及び、該蛋白質に付着されるコード化ポリヌクレオチドを含み、前記蛋白質が、複数の標的のうち所定の標的に特異的に結合し、前記コード化ポリヌクレオチドが、前記アレイに付着される複数のポリヌクレオチドのうち配列特異的且つ位置的に区別可能なポリヌクレオチドに特異的に結合し、各蛋白質及びコード化ポリヌクレオチドが互いから結合的に区別可能である、ポリヌクレオチドによりコード化される蛋白質をさらに含む、請求項7に記載のアレイ。 - 請求項1乃至8のいずれか一項に記載のアレイを含む、マイクロ流体装置。
- 流体試料の流体成分を分離する分離ユニットをさらに含む、請求項9に記載のマイクロ流体装置であって、前記分離ユニットが、
前記注入口と流体連絡し、流れチャネル抵抗を有した流れマイクロ流体チャネル、及び
前記流れチャネルと流体連絡し、アッセイチャネル抵抗を有したアッセイマイクロ流体チャネルを含み、
前記流れマイクロ流体チャネル抵抗及び前記アッセイマイクロ流体チャネル抵抗は、前記流れマイクロ流体チャネルから前記アッセイマイクロ流体チャネルまで前記流体成分の流れを制御するようされ、
前記アレイが前記アッセイマイクロフルイディックス上に配置される、マイクロ流体装置。 - 前記少なくとも1つの標的が複数の標的であり、前記少なくとも1つの捕獲剤又はその構成成分が複数の捕獲剤又はその構成成分であり、前記複数の捕獲剤の各捕獲剤が、互いから結合的に区別可能且つ位置的に区別可能であり、前記複数の捕獲剤の各捕獲剤は、前記複数の標的の各標的に特異的に結合して捕獲剤標的結合複合体を形成する能力を持つ、請求項9又は10に記載のマイクロ流体装置。
- 前記複数の捕獲剤又はその構成成分が、
前記アレイに付着される複数のアレイポリヌクレオチドであり、該アレイに付着される複数のアレイポリヌクレオチドの各ポリヌクレオチドが配列特異的で且つ互いから位置的に区別可能である、アレイポリヌクレオチドを含む、請求項11に記載のマイクロ流体装置。 - 前記複数の捕獲剤又はその構成成分が、
複数のポリヌクレオチドによりコード化される蛋白質であり、各ポリヌクレオチドによりコード化される蛋白質が、蛋白質、及び、該蛋白質に付着されるコード化ポリヌクレオチドを含み、前記蛋白質が、複数の標的のうち所定の標的に特異的に結合し、前記コード化ポリヌクレオチドが、前記アレイに付着される複数のポリヌクレオチドのうち配列特異的且つ位置的に区別可能なポリヌクレオチドに特異的に結合し、各蛋白質及びコード化ポリヌクレオチドが互いから結合的に区別可能である、ポリヌクレオチドによりコード化される蛋白質を含む、請求項12に記載のマイクロ流体装置。 - 試料内の少なくとも1つの標的を検出するためのシステムであって、
請求項1乃至8のいずれか一項に記載のアレイ、及び
該アレイ上のバーコードパターンを検出するための装置、
を含むシステム。 - 前記少なくとも1つの標的は複数の標的であり、前記少なくとも1つの捕獲剤又はその構成成分は複数の捕獲剤又はその構成成分であり、前記複数の捕獲剤の各捕獲剤は、互いから結合的に区別可能且つ位置的に区別可能であり、前記複数の捕獲剤の各捕獲剤は、前記複数の標的の各標的に特異的に結合して捕獲剤標的結合複合体を形成する能力を持つ、請求項14に記載のシステム。
- 前記アレイのバーコードパターンが生物学的プロファイルと付随し、前記バーコードパターンを検出するための装置が前記生物学的プロファイルの可視指標を提供する、請求項15に記載のシステム。
- 前記アレイのバーコードパターンが診断指標と付随し、前記バーコードパターンを検出するための装置が前記診断指標の可視指標を提供する、請求項15に記載のシステム。
- 試料内の複数の標的を検出するためのシステムであって、
請求項7に記載のアレイ、及び
複数のポリヌクレオチドによりコード化される蛋白質であり、各ポリヌクレオチドによりコード化される蛋白質が、蛋白質、及び、該蛋白質に付着されるコード化ポリヌクレオチドを含み、前記蛋白質が、前記複数の標的のうち所定の標的に特異的に結合し、前記コード化ポリヌクレオチドが、前記アレイに付着される複数のポリヌクレオチドのうち配列特異的且つ位置的に区別可能なポリヌクレオチドに特異的に結合し、各蛋白質及びコード化ポリヌクレオチドが互いから結合的に区別可能である、ポリヌクレオチドによりコード化される蛋白質、
を含むシステム。 - 複数のラベルされた分子であって、各ラベルされた分子が前記複数の標的のうち1つの標的に特異的に結合する成分、及び、該成分に付着されるラベル化合物を含み、該ラベル化合物がラベリング信号を提供し、各ラベルされた分子が互いから検出可能な程度に区別可能である、ラベルされた分子をさらに含む、請求項18に記載のシステム。
- 試料内の複数の標的を検出する方法であって、
前記複数の捕獲剤との前記複数の標的の結合を可能にし、捕獲剤標的結合複合体を形成する時間及び状況下で、請求項2乃至5のいずれか一項に記載のアレイに前記試料を接触させるステップ、並びに、
前記捕獲剤標的結合複合体を検出するステップ、
を含む方法。 - 少なくとも1つの検出可能な標的を検出するための基板であって、バーコードパターンを形成する実質的に平行な線に沿った当該基板上への前記少なくとも1つの検出可能な標的の付着を可能にするよう構成される基板。
- 請求項21に記載の基板を含むマイクロ流体装置。
- 複数の検出可能な標的を検出するためのシステムであって、
請求項21に記載の基板、及び
前記バーコードパターンを検出するための装置、
を含むシステム。 - 試料内の複数の標的を検出する方法であって、
前記基板との前記複数の標的の結合を可能にする時間及び状況下で、請求項21に記載の基板に前記試料を接触させるステップ、並びに、
前記基板に付着された前記複数の標的を検出するステップ、
を含む方法。 - 所定のマイクロ流体パターンに沿って分子を流体支持体上に付着させる方法であって、
注入口及び排出口をそれぞれ有する流体チャネルを含んだ骨組を提供するステップであり、前記チャネルの排出口のそれぞれが前記所定のパターンの一部を形成するよう構成されるステップ、
前記骨組と結合するのに適した前記支持体を提供するステップ、
前記骨組みを前記支持体と結合させるステップ、
前記支持体上での前記分子の付着を可能にする時間及び状況下で、前記流体チャネル内に前記分子を提供するステップ;並びに、
前記骨組を前記支持体から離すステップ、
を含む方法。 - 前記パターンが、バーコードパターンを形成する実質的に平行な線を含む、請求項25に記載の方法。
- 前記支持体が、請求項21に記載の基板である、請求項26に記載の方法。
- 前記支持体が、請求項1乃至8のいずれか一項に記載のアレイを含む、請求項26又は27に記載の方法。
- 前記流体支持体がマイクロ流体支持体であり、前記流体チャネルがマイクロ流体チャネルである、請求項25乃至28のいずれか一項に記載の方法。
- 所定のマイクロ流体パターンに沿って分子を流体支持体上に付着させるシステムであって、
注入口及び排出口を有するマイクロ流体チャネルを含んだ骨組であり、前記チャネルの排出口が前記所定のパターンの一部を形成するよう構成される、骨組、並びに、
前記骨組と結合するのに適した支持体、
を含むシステム。 - 前記パターンが、バーコードパターンを形成する実質的に平行な線を含む、請求項30に記載のシステム。
- 前記支持体が、請求項21に記載の基板である、請求項31に記載のシステム。
- 前記支持体が、請求項1乃至8のいずれか一項に記載のアレイを含む、請求項31又は32に記載の方法。
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JP2019528810A (ja) * | 2016-07-07 | 2019-10-17 | ヴァンダービルト ユニヴァーシティ | 疾患材料の検出、捕捉、または除去のための流体デバイス |
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EP2167633A4 (en) | 2014-12-24 |
WO2009012343A2 (en) | 2009-01-22 |
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CA2694545A1 (en) | 2009-01-22 |
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CA2694545C (en) | 2019-10-01 |
EP2167634A4 (en) | 2013-11-06 |
EP3628729A3 (en) | 2020-05-06 |
EP3628729A2 (en) | 2020-04-01 |
JP2010533869A (ja) | 2010-10-28 |
WO2009012340A2 (en) | 2009-01-22 |
AU2008276024A1 (en) | 2009-01-22 |
AU2008276027B2 (en) | 2014-09-04 |
CA2694541A1 (en) | 2009-01-22 |
US20090053732A1 (en) | 2009-02-26 |
US20190195869A1 (en) | 2019-06-27 |
WO2009012343A3 (en) | 2009-04-23 |
AU2008276027A1 (en) | 2009-01-22 |
US10928389B2 (en) | 2021-02-23 |
US20160011189A1 (en) | 2016-01-14 |
WO2009012340A3 (en) | 2009-03-26 |
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