JP2011514522A5 - - Google Patents
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- JP2011514522A5 JP2011514522A5 JP2010547852A JP2010547852A JP2011514522A5 JP 2011514522 A5 JP2011514522 A5 JP 2011514522A5 JP 2010547852 A JP2010547852 A JP 2010547852A JP 2010547852 A JP2010547852 A JP 2010547852A JP 2011514522 A5 JP2011514522 A5 JP 2011514522A5
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Claims (19)
- (a)細胞抽出物を生成するために抗癌剤の投与後、または抗癌剤とのインキュベーション前に単離された乳房腫瘍の細胞を溶解させる工程と;
(b)前記細胞抽出物中の1つ以上の分析物の活性化状態を、前記1つ以上の分析物に対して特異的である複数の希釈系列の捕捉抗体を含むアッセイを用いて検出する工程であって、前記捕捉抗体が固体支持体上に固定され、そして
前記アッセイが、
(i)複数の捕捉分析物を形成するために前記細胞抽出物を捕捉抗体の複数の希釈系列とインキュベートする工程と;
(ii)複数の検出可能な捕捉分析物を形成するために、前記複数の捕捉分析物を、前記複数の活性化状態非依存性抗体および前記対応する分析物に対して特異的である複数の活性化状態依存性抗体を含む検出抗体とインキュベートする工程であって、
前記活性化状態非依存性抗体はグルコースオキシダーゼで標識され、前記グルコースオキシダーゼ及び活性化状態非依存性抗体はスルフヒドリル活性化デキストラン分子へ結合され、前記活性化状態依存性抗体はシグナル増幅対の第1メンバーで標識され、前記グルコースオキシダーゼは前記シグナル増幅対の第1メンバーにチャネリングして反応する酸化剤を生成する工程と、
(iii)増幅シグナルを生成するために、前記複数の検出可能な捕捉分析物をシグナル増幅対の第2メンバーとインキュベートする工程と;
(iv)前記シグナル増幅対の第1および第2メンバーから生成された前記増幅シグナルを検出する工程と;を含む
前記検出工程と;
(c)前記抗癌剤が、前記1つ以上の分析物について検出された活性化状態を前記抗癌剤の非存在下で作成された対照活性化プロファイルと比較することによって、前記乳房腫瘍の治療のために適切であるか、または不適切であるかを決定し、前記方法は乳房腫瘍を治療するのに適切な抗癌剤を選択するためである、工程と、
を含む方法。 - 前記工程(c)に代えて、
前記1つ以上の分析物について検出された活性化状態を前記抗癌剤の非存在下で作成された対照活性化プロファイルと比較することによって、前記乳房腫瘍が前記抗癌剤を用いた治療に応答性または非応答性であると同定し、前記方法が抗癌剤を用いた治療への乳房腫瘍の応答を同定するための方法である、工程を含む請求項1に記載の方法。 - 前記工程(c)に代えて、
前記1つ以上の分析物について検出された活性化状態を前記抗癌剤の非存在下で作成された対照活性化プロファイルと比較することによって、前記被験者が前記抗癌剤を用いた治療に応答する可能性を予測し、前記方法が抗癌剤を用いた治療への乳房腫瘍を有する被験者の応答を予測するための方法である、工程を含む請求項1に記載の方法。 - 前記乳房腫瘍は、乳管癌を有する被験者に由来し、前記乳管癌は好ましくは浸潤性乳管癌または非浸潤性乳管癌であるか、又は前記乳房腫瘍は、小葉癌を有する被験者に由来し、前記小葉癌は、好ましくは浸潤性小葉癌または上皮内小葉癌である、請求項1〜3のいずれか一項に記載の方法。
- 前記細胞は、前記乳房腫瘍の循環細胞を含み、前記循環細胞は、好ましくは免疫磁気分離法によってサンプルから単離され、前記サンプルは、好ましくは全血、血清、血漿、管洗浄、乳頭吸引液、リンパ液、骨髄吸引液、尿、唾液、細針吸引液、およびそれらの組み合わせからなる群より選択され、前記循環細胞は、好ましくは循環腫瘍細胞、循環内皮細胞、循環内皮前駆細胞、癌幹細胞、播種性腫瘍細胞、およびそれらの組み合わせからなる群より選択される、請求項1〜3のいずれか一項に記載の方法。
- 前記細胞は、腫瘍組織から単離され、前記腫瘍組織は、好ましくは原発性腫瘍組織または転移性腫瘍組織であり、前記細胞は、好ましくは細針吸引液サンプルとして腫瘍組織から単離される、請求項1〜3のいずれか一項に記載の方法。
- 前記単離した細胞は、増殖因子を用いてインビトロ(in vitro)で刺激される、請求項1〜3のいずれか一項に記載の方法。
- 前記抗癌剤は、モノクローナル抗体、チロシンキナーゼ阻害剤、化学療法薬、ホルモン療法薬、放射線療法薬、ワクチン、およびそれらの組み合わせからなる群より選択され、
前記モノクローナル抗体は、好ましくはトラスツズマブ(Herceptin(登録商標))、アレムツズマブ(Campath(登録商標))、ベバシズマブ(Avastin(登録商標))、セツキシマブ(Erbitux(登録商標))、ゲムツズマブ(Mylotarg(登録商標))、パニツムマブ(Vectibix(商品名))、リツキシマブ(Rituxan(登録商標))、トシツモマブ(BEXXAR(登録商標))、およびそれらの組み合わせからなる群より選択され、
前記チロシンキナーゼ阻害剤は、好ましくはゲフィチニブ(Iressa(登録商標))、スニチニブ(Sutent(登録商標))、エルロチニブ(Tarceva(登録商標))、ラパチニブ(Tykerb(登録商標))、カネルチニブ(CI1033)、セマキシニブ(SU5416)、バタラニブ(PTK787/ZK222584)、ソラフェニブ(BAY43−9006)、イマチニブメシレート(Gleevec(登録商標))、レフルノミド(SU101)、バンデタニブ(ZACTIMA(商品名);ZD6474)、およびそれらの組み合わせからなる群より選択され、
前記化学療法薬は、好ましくはペメトレキセド(ALIMTA(登録商標))、ゲムシタビン(Gemzar(登録商標))、シロリムス(ラパマイシン)、ラパマイシンアナログ、白金化合物、カルボプラチン、シスプラチン、サトラプラチン、パクリタキセル(Taxol(登録商標))、ドセタキセル(Taxotere(登録商標))、テムシロリムス(CCI−779)、エベロリムス(RAD001)、およびそれらの組み合わせからなる群より選択され、
前記ホルモン療法薬は、好ましくはアロマターゼ阻害剤、選択的エストロゲン受容体調節剤、ステロイド、フィナステリド、ゴナドトロピン放出ホルモンアゴニスト、それらの薬学的に許容される塩、それらの立体異性体、それらの誘導体、それらのアナログ、およびそれらの組み合わせからなる群より選択され、
前記放射線療法薬は、好ましくは 47 Sc、 64 Cu、 67 Cu、 89 Sr、 86 Y、 87 Y、 90 Y、 105 Rh、 111 Ag、 111 In、 117m Sn、 149 Pm、 153 Sm、 166 Ho、 177 Lu、 186 Re、 188 Re、 211 At、および 212 Bi、およびそれらの組み合わせからなる群より選択される、
請求項1〜3のいずれか一項に記載の方法。 - 前記1つ以上の分析物は、複数のシグナル伝達分子を含み、
前記複数のシグナル伝達分子は、好ましくは受容体チロシンキナーゼ、非受容体チロシンキナーゼ、チロシンキナーゼシグナル伝達カスケード成分、核ホルモン受容体、核受容体コアクチベーター、核受容体リプレッサー、およびそれらの組み合わせからなる群より選択され、
前記複数のシグナル伝達分子は、好ましくはEGFR(ErbB1)、HER−2(ErbB2)、p95ErbB2、HER−3(ErbB3)、HER−4(ErbB4)、Raf、SRC、Mek、NFkB−IkB、mTor、PI3K、VEGF、VEGFR−1、VEGFR−2、VEGFR−3、Eph−a、Eph−b、Eph−c、Eph−d、cMet、FGFR、cKit、Flt−3、Tie−1、Tie−2、Flt−3、cFMS、PDGFRA、PDGFRB、Abl、FTL3、RET、Kit、HGFR、FGFR1、FGFR2、FGFR3、FGFR4、IGF−1R、ER、PR、NCOR、AIB1、およびそれらの組み合わせからなる群より選択され、又は
前記複数のシグナル伝達分子は、好ましくはErbB1、ErbB2、p95ErbB2、ErbB3、ErbB4、VEGFR−1、VEGFR−2、VEGFR−3、ER、PR、およびそれらの組み合わせからなる群より選択される、
請求項1〜3のいずれか一項に記載の方法。 - 前記活性化状態は、リン酸化状態、ユビキチン化状態、複合体形成状態、およびそれらの組み合わせからなる群より選択される、請求項1〜3のいずれか一項に記載の方法。
- 前記固体支持体は、ガラス、プラスチック、チップ、ピン、フィルター、ビーズ、紙、膜、繊維束、およびそれらの組み合わせからなる群より選択される、請求項1〜3のいずれか一項に記載の方法。
- 前記捕捉抗体は、アドレス可能なアレイ内の固体支持体上に固定される、請求項1〜3のいずれか一項に記載の方法。
- 前記活性化状態依存性抗体は、前記シグナル増幅対の第1メンバーを用いて直接的に標識されるか、又は
前記活性化状態依存性抗体は、前記シグナル増幅対の第1メンバーを用いて、前記活性化状態依存性抗体に結合された結合対の第1メンバーと前記シグナル増幅対の第1メンバーに結合された前記結合対の第2メンバーとの間の結合によって標識され、
前記結合対の第1メンバーは、好ましくはビオチンであり、
前記結合対の第2メンバーは、好ましくはストレプトアビジンである、請求項1〜3のいずれか一項に記載の方法。 - 前記スルフヒドリル活性化デキストラン分子は、500kDaの分子量を有する、請求項1〜3のいずれか一項に記載の方法。
- 前記酸化剤は、過酸化水素(H2O2)である、請求項1〜3のいずれか一項に記載の方法。
- 前記シグナル増幅対の第1メンバーは、ペルオキシダーゼ、好ましくは西洋ワサビペルオキシダーゼ(HRP)であり、及び/又は前記シグナル増幅対の第2メンバーは、チラミド試薬、好ましくはビオチン−チラミドである、請求項15に記載の方法。
- 前記増幅シグナルは、活性化チラミドを生成するために前記ビオチン−チラミドのペルオキシダーゼ酸化によって生成される、請求項16に記載の方法。
- 前記活性化チラミドは、直接検出されるか、又は前記活性化チラミドは、シグナル検出試薬の添加に基づいて検出され、前記シグナル検出試薬は、好ましくはストレプトアビジン標識フルオロフォアであるか、又はストレプトアビジン標識ペルオキシダーゼおよび発色試薬の組み合わせであり、前記発色試薬は、好ましくは3,3’,5,5’−テトラメチルベンジジン(TMB)である、請求項17に記載の方法。
- 固体支持体上に固定された複数の希釈系列の捕捉抗体を含む優れたダイナミックレンジを有するアレイであって、各希釈系列中の捕捉抗体は細胞抽出物中のシグナル伝達経路の成分に対応する1つ以上の分析物に対して特異的である、アレイの請求項1〜3のいずれか一項に記載の方法を行うための使用。
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JP2008292424A (ja) | 2007-05-28 | 2008-12-04 | Nippon Medical School | 腫瘍の検出方法 |
ES2526211T3 (es) * | 2007-07-13 | 2015-01-08 | Nestec S.A. | Selección de fármacos para la terapia del cáncer de pulmón utilizando matrices basadas en anticuerpos |
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KR101851425B1 (ko) | 2011-09-02 | 2018-04-23 | 네스텍 소시에테아노님 | 치료 효능 결정을 위한 신호 경로 단백질의 프로파일링 |
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