JP2009532032A - 上部に配置された局在化分子を有する物品およびそれを作製する方法 - Google Patents
上部に配置された局在化分子を有する物品およびそれを作製する方法 Download PDFInfo
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Abstract
Description
本出願は、デイビッド R.ランク(David R.Rank)らによる「ARTICLES HAVING LOCALIZED MOLECULES DISPOSED THEREON AND METHODS OF PRODUCING SAME」という表題の2006年3月30日出願の米国仮特許出願第11/394,352号明細書の一部継続出願であり、それはあらゆる目的でその全体が参照により本明細書中に援用される。
本発明の一部は、国立ヒトゲノム研究所(NHGRI)助成金番号R01−HG003710−01の下で政府の助成によって行われたものであり、政府は本発明に対する特定の権利を有しうる。
図1は用途におけるゼロモード導波路(ZMW)の概略図を示す。
本発明は、一般に、所望の分子を基板上の予備選択された位置もしくはエリア内にまたは規定の体積内に提供するための方法およびプロセス、ならびにかかる方法またはプロセスから、特に所望の低濃度でまたは個々の分子として、光閉じ込め内で作られる物品を対象とする。特に好ましい態様では、本発明は、特定の空間または体積内に個々の分子を、分子の空間的個別性が例えば化学的、光学的、電気的、またはそれらに類する原理で用いられうるように局在化するための方法を対象とする。本発明は、これらのプロセスにより作製される基板、デバイス、容器など、例えば光閉じ込めも提供する。本発明のプロセスが種々の所与の所望の空間または体積のタイプのいずれかの範囲内に個々の分子を提供するのに広範に実用的でありうる一方、特に好ましい態様では、同プロセスを用い、所望の分子、例えば酵素が光閉じ込めの光学的に接近可能な部分、特にゼロモード導波路(ZMW)の内部に選択的に堆積または固定化される。
上で示されるように、少なくとも1つの態様では、加法的プロセスを用い、本発明の所望の固定化分子が提供される。加法的プロセスは、典型的には所望の位置での結合基または連結基の選択的供給と、その後の目的の分子の堆積に依存する。さらに、この堆積は加法的または減法的プロセスの結果でありうる。
のエステルを含む。
の
および
(各々、ローディア(Rhodia)からAquarite(登録商標)EC4020およびAquarite(登録商標)ESLとして市販)
など、末端キャップされた(end−capped)ホスホン酸塩、ならびに
ビニルホスホン酸−アクリル酸共重合体
(ローディア(Rhodia)からAlbritect(商標)CP30として市販)
などの共重合体などの高分子量の高分子ホスホン酸塩を含む。
上記のように、他の態様では、減法的プロセスを用いることで目的の分子が基板の所望領域内にかつ分子の所望の濃度および/または密度で提供される。上記のように、減法的プロセスは一般に特徴づけられており、目的の分子を例えば所望領域内を含む基板表面全体にわたりより遍在的に堆積させる点で上記の加法的プロセスとは異なる。次いで、例えば非所望領域内に位置する過剰な目的の分子が除去される。かかる減法的プロセスでは、種々の異なるプロセスが用いられうる。
ガラス基板層の上に堆積されたアルミニウムクラッド層を有するガラス層を含む基板が使用可能である。ZMWコアのアレイをクラッド層内に作製し、クラッド層を通してガラス基板に対して開口を提供する。場合により基板全体をさらに処理し、例えばクラッド層およびコアの上部に薄い絶縁層を提供し、実質的に均一の表面を提供する。かかる層は、典型的には、例えばCVDおよびMVD法を含む蒸着技術、ならびに例えばスピンオンガラス系を用いる流体堆積または原位置形成などの他の方法により適用されるSiO2コーティングを含む。基板表面を誘導体化し、まず表面に連結されたアミノ末端基の相対的に均一な集団を提供する。例えばガラス表面においては、かかる誘導体化では典型的に当該技術分野で既知の標準のアミノシランの化学反応を用いる。あるいは、アミン基を、既存の水酸基を介して表面かまたはそれ以外では誘導体化された表面に連結されたリンカー分子上に提供可能である。かかる連結基を、最終的に表面に結合されることになる目的の分子の密度をさらに制御するため、制限された密度で提供可能である(例えば、2005年9月30日出願の同一出願人による米国特許出願第11/240,662号明細書(あらゆる目的でその全体が参照により本明細書中に援用される)を参照)。
(プライミング層を提供するため)PDMSガスケットの存在下で予めプラズマ処理したZMWに、クラッド層の上面上を含むアレイの表面全体にわたり実質的に均一に表面吸着された(環状鋳型核酸と複合体を形成した)Φ29N62DDNAポリメラーゼを提供した。
(シグマ・ケミカル社(Sigma Chemical Co.)、P0803またはP9290)と、25mMトリス−HCl、pH7.5、10mM β−メルカプトエタノール、1mM EDTAの中、室温で5分間接触させた。ビーズ直径はアレイ上の導波路コアの公称直径よりもはるかに大きく、ビーズまたはそれと結合したプロテアーゼ分子のコアへの侵入がいかなる実質的程度であっても阻止された。
以下に、ZMWの底面上でのDNAポリメラーゼの選択的固定化および高分子電解質多層を有する残りのZMW表面の不動態化を実証する一連の実験を示す。シランとガラスおよび酸化アルミニウムとの異なる反応性を用いるプロセスを図18に概略的に図示する。PEG−ビオチンのシラン化処理が用いられる条件下でガラスに特異的であることにより、ZMW底面のみに化学的誘導体化がもたらされる。次いで、アルミニウム層を高分子電解質多層、この実施例ではPAA/PEI/PAA/PEI/PAA(ここでPAAはポリ(アクリル酸)、PEIはポリ(エチレンイミン)である)の2.5倍の多層を用いて不動態化する。ビオチンでタグ化されたポリメラーゼは高分子電解質多層により拒絶されるが、アビジン化学反応を介してビオチン化PEG表面に結合し、それによりポリメラーゼのZMWの底面へのバイアスされた固定化がもたらされる。さらに、高分子電解質多層はヌクレオチド類似体のアルミニウム層への非特異的結合を制限する。
ポリビニルホスホン酸(PVPA)を未処理のZMW上に堆積する結果、アルミニウム表面に結合する非特異的タンパク質(例えばニュートラアビジンおよびポリメラーゼ)およびヌクレオチド類似体から不動態化されたZMWが得られる。PVPAは、アルミニウムに特異的であり、ZMWのSiO2底面に作用することなく、非特異的な捕捉剤あるいは特異的なポリメラーゼ堆積のための重合固定化またはその後の誘導体化(例えばPLL−PEGなどの化合物のシラン化処理または結合による)において用いられうる。
Claims (87)
- 選択的に活性な領域を上部に有する基板を作製する方法であって、
上部に画成された複数のナノ構造を有する基板を提供するステップであって、ここで前記ナノ構造の各々によって、第1の活性化制御物質の、前記基板の表面の選択された部分上に活性な化学基を提供する能力が制限される、ステップと、
前記基板の前記表面の前記選択された部分上に活性な化学基を選択的に提供するように、前記基板を少なくとも前記第1の活性化制御物質に暴露するステップと
を含む方法。 - 前記ナノ構造によって、前記基板の前記表面の前記選択された部分が前記第1の活性化制御物質へ暴露されることが実質的に阻止される、請求項1に記載の方法。
- 前記提供するステップが、前記基板の前記表面上に非活性化可能な化学官能基を提供するステップを含み、かつ前記第1の活性化制御物質が、前記基板の前記表面の前記選択された部分以外の部分上で前記化学官能基を非活性化する、請求項2に記載の方法。
- 前記ナノ構造によって、前記基板表面の前記選択された部分以外の部分が前記第1の活性化制御物質へ暴露されることが実質的に阻止される、請求項1に記載の方法。
- 前記提供するステップが、前記基板の前記表面上に活性化可能な化学官能基を提供するステップを含み、かつ前記第1の活性化制御物質が、前記基板の前記表面の前記選択された部分上で前記化学官能基を活性化する、請求項4に記載の方法。
- 前記ナノ構造が、前記基板の前記表面内に配置されたナノスケールのウェルを含む、請求項1に記載の方法。
- 前記ナノ構造が、それ以外の部分は不透明な基板内に配置されたナノスケールの光開口部を含む、請求項1に記載の方法。
- 前記提供するステップが、前記基板の前記表面上に化学官能基を提供するステップを含み、前記化学官能基が、前記活性化制御物質へ暴露されると活性化または非活性化される、請求項1に記載の方法。
- 前記ナノ構造がナノスケールのウェルを含み、ここで前記選択された領域が前記ナノスケールのウェルの底面を含み、
前記活性化制御物質が、化学官能基を同伴しかつ前記ナノスケールのウェル内に実質的に拡散することのない同伴マトリックスを含み、かつ、
前記同伴マトリックスを除去し、化学官能基を前記基板の前記表面の前記選択された領域内以外の部分から除去するステップをさらに含む、
請求項1に記載の方法。 - 前記第1の活性化制御物質が、前記基板の前記表面上の化学官能基を活性化または非活性化するように選択された波長の光を含む、請求項1に記載の方法。
- 前記光が前記化学官能基を活性化するように選択され、かつ前記ナノ構造によって、前記基板表面の前記選択された部分以外の部分が前記第1の活性化制御物質へ暴露されることが実質的に阻止される、請求項10に記載の方法。
- 前記化学官能基が、ニトロベラトリル、1−ピレニルメチル、6−ニトロベラトリルオキシカルボニル、ジメチルジメトキシベンジルオキシカルボニル、2−ニトロベンジルオキシカルボニル、メチル、メチル−6−ニトロピペロニルオキシカルボニル、2−オキシメチレンアントラキノン、ジメトキシベンジルオキシカルボニル、5−ブロモ−7−ニトロインドリニル、o−ヒドロキシ−α−メチルシンナモイル、およびそれらの混合物から選択される光活性化可能な基を含む、請求項10に記載の方法。
- 前記光が前記化学官能基を非活性化するように選択され、かつ前記ナノ構造によって、前記基板の前記表面の前記選択された部分が前記第1の活性化制御物質へ暴露されることが実質的に阻止される、請求項10に記載の方法。
- 前記化学官能基が、光切断可能なリンキング基を介して前記基板の前記表面に連結される、請求項13に記載の方法。
- 前記化学官能基が第1の活性化可能な化学官能基を含み、
前記第1の活性化制御物質が、前記第1の活性化可能な化学官能基を活性化するように選択された波長の光を含み、
前記基板が不透明層および透明層を含み、
前記ナノ構造が、前記不透明層を通して前記透明層に対して配置されたゼロモード導波路を含み、かつ、
前記暴露するステップが、前記光を前記基板の前記透明層に誘導し、前記第1の活性化可能な化学官能基を活性化し、前記透明層の上または近傍にある前記ゼロモード導波路内に第1の活性な化学官能基を提供するステップを含む、
請求項10に記載の方法。 - 前記暴露するステップで提供される前記第1の活性な化学官能基をキャップし、前記暴露するステップでの前記光とは異なる波長の光により活性化可能な第2の活性化可能な化学官能基を提供するステップと、
前記第1の活性化可能な化学官能基ではなく前記第2の活性化可能な化学官能基を活性化するように選択された波長の光を前記基板の前記透明層に誘導するステップをさらに含み、
前記誘導するステップが、前記第1の暴露するステップで活性化された前記化学官能基の一部を活性化するための条件下で実施される、
請求項15に記載の方法。 - 前記化学官能基が非活性化可能な化学官能基を含み、
前記第1の活性化制御物質が、前記非活性化可能な化学官能基を非活性化するように選択された波長の光を含み、
前記基板が不透明層および透明層を含み、
前記ナノ構造が、前記不透明層を通して前記透明層に対して配置されたゼロモード導波路を含み、かつ、
前記暴露するステップが前記基板の前記不透明層に前記光を誘導し、前記不透明層上で化学官能基を非活性化するステップを含む、
請求項10に記載の方法。 - 前記非活性化可能な化学官能基が、光切断可能なリンキング基を介して前記基板の前記表面に連結された活性な化学官能基を含む、請求項17に記載の方法。
- 前記非活性化可能な化学官能基が、前記暴露するステップにおける前記光への暴露により少なくとも部分的に非活性化されるタンパク質を含む、請求項17に記載の方法。
- 前記非活性化可能な化学官能基がフルオロフォア結合タンパク質を含み、かつ前記暴露するステップが前記フルオロフォアの存在下で実施される、請求項19に記載の方法。
- 前記フルオロフォア結合タンパク質が核酸ポリメラーゼを含む、請求項20に記載の方法。
- 前記第1の活性化制御物質が、前記化学官能基を活性化または非活性化する化学剤を含む、請求項1に記載の方法。
- 前記第1の活性化制御物質が、前記化学官能基を非活性化する化学剤を含み、
前記ナノ構造が、前記化学剤のナノスケールのウェルへの拡散を制限する、前記基板の前記表面内に画成された前記ナノスケールのウェルを含み、かつ
前記暴露するステップが、前記ナノスケールのウェルの底面またはその近傍の表面の部分以外の前記基板の前記表面上の化学官能基を非活性化する条件下で、前記基板を前記化学剤に暴露するステップを含む、請求項22に記載の方法。 - 前記化学剤が、化学官能基に結合し、それらを不活性化するキャッピング基を含む、請求項22に記載の方法。
- 前記化学剤が、前記ナノスケールのウェルへ入ることから少なくとも部分的に排除される排除成分に連結される、請求項22に記載の方法。
- 前記排除成分が大粒子および巨大分子から選択される、請求項25に記載の方法。
- 前記粒子がビーズを含む、請求項26に記載の方法。
- 前記化学剤が非活性化成分を含む、請求項26に記載の方法。
- 前記非活性化成分が酵素を含む、請求項28に記載の方法。
- 前記酵素がプロテアーゼ、ヌクレアーゼ、およびカルボヒドラーゼから選択される、請求項29に記載の方法。
- 選択された活性領域を上部に有する分析用基板を製造する方法であって、
基板の選択された領域に対する改良された光学的アクセスをもたらす、上部に配置された光学的分析構造を有する基板を提供するステップと、
第1の電磁放射により活性化または非活性化可能な表面官能基を提供するステップと、
前記基板に前記第1の電磁放射を誘導するステップであって、それにより前記光学的分析構造が前記電磁放射を誘導し、前記基板の前記選択された領域内で前記表面官能基を選択的に活性化または非活性化し、前記基板の活性領域を選択的に提供する、ステップと
を含む、方法。 - 実質的に導波路の底部に活性な化学的表面を有するゼロモード導波路を提供する方法であって、
基板内に配置されたゼロモード導波路を提供するステップと、
前記ゼロモード導波路の表面上に化学官能基を提供するステップと、
前記ゼロモード導波路の前記表面の第2の部分ではなく第1の部分を、前記第1の部分上の前記化学官能基を選択的に活性化または非活性化する活性化制御物質に暴露し、実質的に前記ゼロモード導波路の底部に活性な化学的表面を提供するステップと、
を含む、方法。 - ゼロモード導波路アレイであって、
クラッド層内に配置された複数のゼロモード導波路コアであって、各コアが底面を有するコアと、
実質的に前記コア内に限られる化学的に活性な表面と、
を含む、ゼロモード導波路アレイ。 - 前記化学的に活性な表面が前記コアの前記底面上に優先的に局在化される、請求項33に記載のアレイ。
- 上部に配置された選択された活性領域を有する基板を提供する方法であって、
上部に画成された光学的改良構造を有する基板を提供するステップであって、ここで前記光学的改良構造は電磁放射を誘導し、前記基板表面の選択された領域の近傍に捕捉力を生成するのに十分な増強された電磁界を、前記基板表面の選択された領域の近傍に提供することが可能である、ステップと、
前記基板において電磁放射を誘導するステップであって、前記選択された領域の近傍の活性分子に対して捕捉力を生成するのに十分な前記増強された電磁界を、前記選択された領域に提供する、ステップと、
前記活性分子を前記選択された領域に連結するステップと、
を含む、方法。 - 上部に配置された選択された活性領域を有する基板を作製する方法であって、
化学官能基からなる表面を有する基板およびその上に画成された複数の分離したナノスケールの反応領域を提供するステップと、
1つもしくは複数の化学官能基または活性化制御物質を選択された領域内の前記表面上にパターニングし、実質的に前記分離したナノスケールの反応領域内に限られる活性のある化学的な機能領域を提供するステップと、
を含む、方法。 - 前記パターニングするステップが、活性化制御物質をマスクを介して誘導し、活性な化学官能基を、実質的に前記分離したナノスケールの反応領域内に限って生成するステップを含む、請求項36に記載の方法。
- 前記活性化制御物質が、前記基板の前記表面上の化学官能基を活性化または非活性化するように選択された波長の光を含む、請求項37に記載の方法。
- 前記パターニングするステップが、前記活性化制御物質をプリンティングプロセスを介して適用するステップを含む、請求項36に記載の方法。
- 核酸分子の配列を同定する方法であって、
基板上の分離した観察領域内に複数の核酸ポリメラーゼ/鋳型/プライマー複合体を提供するステップと、鋳型依存性の方法でヌクレオチドまたはヌクレオチド類似体の連続的付加を検出し、前記複数の観察領域内での前記ヌクレオチドまたはヌクレオチド類似体の取り込まれた配列を同定するステップと、を含み、
ここで、前記基板が前記観察領域外部の領域内でポリメラーゼ活性、ポリメラーゼの存在、鋳型の存在およびプライマーの存在のうちの1つもしくは複数を実質的に低減するように製造されている、方法。 - 核酸分子の配列を同定する方法であって、
基板表面上の分離した観察領域内に複数の核酸ポリメラーゼ/鋳型/プライマー複合体を提供するステップと、鋳型依存性の方法でヌクレオチドまたはヌクレオチド類似体の連続的付加を検出し、前記複数の観察領域内での前記ヌクレオチドまたはヌクレオチド類似体の取り込まれた配列を同定するステップと、
少なくとも第1および第2の分離した観察領域の間の前記基板表面上に観察領域内バリアを提供し、1つもしくは複数の反応物または生成物の観察領域内での拡散を実質的に阻止するステップと、
を含む、方法。 - 前記バリアが、第1の流体と、前記第1の観察領域からの1つもしくは複数の生成物または反応物の前記第2の観察領域への到達を実質的に阻止するのに十分な前記第1および第2の観察領域の間の距離とを含む、請求項41に記載の方法。
- 前記基板表面が平面状である、請求項42に記載の方法。
- 前記基板表面が構造化される、請求項42に記載の方法。
- 前記バリアが、前記基板表面内に配置された少なくとも第1および第2のナノスケールのウェルをさらに含み、ここで前記第1および第2の観察領域がそれぞれ、前記第1および第2のナノスケールのウェル内に配置される、請求項44に記載の方法。
- 基板上に配置される光閉じ込め内に所望の分子を優先的に局在化する方法であって、
前記所望の分子を前記基板の前記表面上部に堆積させるステップと、
前記所望の分子を前記光閉じ込め内でない前記基板の前記表面から選択的に除去するステップと、
を含む、方法。 - 前記基板が不透明層および透明層を含み、かつここで前記光閉じ込めが、前記不透明層を通して前記透明層に対して配置されたゼロモード導波路を含む、請求項46に記載の方法。
- 前記所望の分子を前記光閉じ込め内でない前記基板の前記表面から選択的に除去するステップが、前記基板を、前記光閉じ込め内に入ることから少なくとも部分的に排除される排除成分に連結された非活性化成分と接触させるステップを含む、請求項46に記載の方法。
- 前記非活性化成分が酵素を含む、請求項48に記載の方法。
- 前記酵素がプロテアーゼ、ヌクレアーゼ、およびカルボヒドラーゼからなる群から選択される、請求項49に記載の方法。
- 前記酵素が部位特異的プロテアーゼである、請求項50に記載の方法。
- 前記排除成分が大粒子または巨大分子である、請求項48に記載の方法。
- 前記排除成分がビーズを含む、請求項52に記載の方法。
- 前記排除成分が硬質もしくは半硬質の細長い高分子を含む、請求項52に記載の方法。
- 前記排除成分が二本鎖核酸分子を含む、請求項52に記載の方法。
- 前記非活性化成分がプロテアーゼであり、かつ前記排除成分が二本鎖DNA分子である、請求項48に記載の方法。
- 前記所望の分子が酵素である、請求項46に記載の方法。
- 前記所望の分子が結合部分を含む、請求項46に記載の方法。
- 前記所望の分子がビオチン分子である、請求項58に記載の方法。
- 基板上に配置される光閉じ込め内に分子を局在化する方法であって、
前記光閉じ込め内部を含む前記基板の前記表面に光活性化された連結基を提供するステップと、
前記基板に活性化放射を誘導するステップであって、ここで前記光閉じ込めによって、活性化放射が前記光閉じ込め内部にのみ入射される、ステップと、
前記分子を前記光活性化された連結基に連結させるステップと、
を含む、方法。 - 目的の分子を基板上に選択的に固定化する方法であって、
異なる表面特性を有する第1の表面成分および第2の表面成分を有する基板を提供するステップと、
前記第1の表面成分の前記表面特性と前記第2の表面成分の前記表面特性の間の差異に基づき、前記目的の分子を前記第1の表面成分に選択的に連結させるステップと、
を含む、方法。 - 前記第1の表面成分がSiO2を含み、かつ前記第2の表面成分が金属もしくは金属酸化物を含む、請求項61に記載の方法。
- 前記異なる表面特性が表面電荷を含む、請求項61に記載の方法。
- 前記第1の表面成分が負の表面電荷を有し、かつ前記第2の表面成分が正の表面電荷を有する、請求項63に記載の方法。
- 前記基板が前記第1の表面成分層上に前記第2の表面成分層を含み、かつここで前記基板は、前記第2の表面成分層を通して前記第1の表面成分層に対して配置されたゼロモード導波路を含む、請求項61に記載の方法。
- 前記目的の分子が前記第1の表面成分に優先的に結合する、請求項61に記載の方法。
- 前記基板が、前記第1および第2の表面成分の前記表面特性の間の前記差異に基づき、前記第1の表面成分と選択的に結合する第1の組成物と接触される、請求項61に記載の方法。
- 前記第1の組成物が第1の連結基を含み、かつここで前記目的の分子を前記第1の表面成分に選択的に連結させるステップが、前記目的の分子を前記第1の連結基に連結させるステップを含む、請求項67に記載の方法。
- 前記第1の連結基がビオチンを含む、請求項68に記載の方法。
- 前記第1の組成物がシランを含む、請求項67に記載の方法。
- 前記第1の組成物がビオチン−PEG−シランを含む、請求項70に記載の方法。
- 前記第1の組成物がリン脂質を含む、請求項67に記載の方法。
- 前記第1の組成物がポリ(L−リジン)−ポリ(エチレングリコール)またはポリ(L−リジン)−ポリ(エチレングリコール)−ビオチンを含む、請求項67に記載の方法。
- 前記基板を、前記第1および第2の表面成分の前記表面特性の間の前記差異に基づき、前記第2の表面成分に選択的に結合する第2の組成物と接触させるステップを含む、請求項61に記載の方法。
- 前記第2の組成物が高分子電解質を含む、請求項74に記載の方法。
- 前記第2の組成物が高分子電解質−PEG共重合体を含む、請求項74に記載の方法。
- 前記第2の表面成分上に高分子電解質多層を堆積させるステップを含む、請求項74に記載の方法。
- 前記第2の組成物が、1つもしくは複数のホスホン酸基を含む化合物を含む、請求項74に記載の方法。
- 前記第2の組成物がポリビニルホスホン酸である、請求項78に記載の方法。
- 前記第2の組成物が、
および
からなる群から選択される、請求項78に記載の方法。 - 前記第2の組成物が、2−カルボキシエチルホスホン酸;アミノトリ(メチレンホスホン酸);1−ヒドロキシエチリデン−1,1,−ジホスホン酸;ヘキサメチレンジアミンテトラ(メチレンホスホン酸);ジエチレントリアミンペンタ(メチレンホスホン酸);エチレンジアミンテトラ(メチレンホスホン酸);ビス(ヘキサメチレントリアミンペンタ(メチレンホスホン酸));2−ホスホノブタン−1,2,4−トリカルボキシル酸;およびモノエタノールアミンジホスホン酸からなる群から選択される、請求項78に記載の方法。
- 前記第2の組成物が、1つもしくは複数のリン酸基を含む化合物を含む、請求項74に記載の方法。
- 前記第2の組成物がアルキルリン酸塩またはアルキルホスホン酸塩を含む、請求項74に記載の方法。
- 前記第2の組成物が、オクチルホスホン酸、デシルホスホン酸、ドデシルホスホン酸、ヘキサデシルホスホン酸、オクタデシルホスホン酸、ドコシルホスホン酸、ヒドロキシ−ドデシルホスホン酸、ヒドロキシ−ウンデセニル−ホスホン酸、デカンジイルビス(ホスホン酸)、ドデシルリン酸、およびヒドロキシ−ドデシルリン酸からなる群から選択される、請求項74に記載の方法。
- 前記第2の組成物が第2の連結基を含む、請求項74に記載の方法。
- 前記目的の分子を前記第1の表面成分に選択的に連結させるステップが、前記基板を前記第1の表面成分に選択的に結合する第1の組成物と接触させるステップと、前記目的の分子を前記第1の組成物に連結させるステップと、を含み、
前記方法が前記基板を前記第2の表面成分に選択的に結合する第2の組成物と接触させるステップをさらに含む、請求項61に記載の方法。 - 目的の分子を基板の選択された領域上に選択的に堆積する方法であって、
導電材料を含む第1の成分および絶縁体を含む第2の成分を有する基板を提供するステップと、
目的の分子と前記第1の成分の表面との結合を増大または低下させるように電位を前記第1の成分に印加するステップと、
を含む、方法。
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JP2011525111A (ja) * | 2008-06-16 | 2011-09-15 | ピーエルシー ダイアグノスティクス, インコーポレイテッド | フェーズ合成による核酸配列決定システム及び方法 |
KR101789586B1 (ko) | 2010-12-06 | 2017-10-26 | 삼성디스플레이 주식회사 | 광 산란 기판, 이의 제조 방법, 이를 포함하는 유기 발광 표시 장치 및 유기 발광 표시 장치의 제조 방법 |
JP2020526774A (ja) * | 2017-07-07 | 2020-08-31 | アヴィアーナ モレキュラー テクノロジーズ,エルエルシー | 弾性表面波センサー用の生物活性コーティング |
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EP2002019A2 (en) | 2008-12-17 |
US20140315756A1 (en) | 2014-10-23 |
US20070238679A1 (en) | 2007-10-11 |
CN102212515B (zh) | 2014-01-08 |
US20080176769A1 (en) | 2008-07-24 |
US20220098660A1 (en) | 2022-03-31 |
WO2007123763A3 (en) | 2008-11-06 |
CA2645758A1 (en) | 2007-11-01 |
US8802600B2 (en) | 2014-08-12 |
US11186871B2 (en) | 2021-11-30 |
US20080032301A1 (en) | 2008-02-07 |
US8772202B2 (en) | 2014-07-08 |
WO2007123763A2 (en) | 2007-11-01 |
US20080153100A1 (en) | 2008-06-26 |
US9944980B2 (en) | 2018-04-17 |
US8975216B2 (en) | 2015-03-10 |
US10655172B2 (en) | 2020-05-19 |
US20200377940A1 (en) | 2020-12-03 |
EP2002019A4 (en) | 2009-07-15 |
ES2385015T3 (es) | 2012-07-17 |
ATE546522T1 (de) | 2012-03-15 |
EP2002019B1 (en) | 2012-02-22 |
CN101426910A (zh) | 2009-05-06 |
AU2007241049B2 (en) | 2012-07-05 |
CA2645758C (en) | 2016-08-09 |
US8193123B2 (en) | 2012-06-05 |
CN102212515A (zh) | 2011-10-12 |
US20120302459A1 (en) | 2012-11-29 |
AU2007241049A1 (en) | 2007-11-01 |
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