JP5937586B2 - 分子間相互作用の検出方法 - Google Patents
分子間相互作用の検出方法 Download PDFInfo
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- JP5937586B2 JP5937586B2 JP2013517458A JP2013517458A JP5937586B2 JP 5937586 B2 JP5937586 B2 JP 5937586B2 JP 2013517458 A JP2013517458 A JP 2013517458A JP 2013517458 A JP2013517458 A JP 2013517458A JP 5937586 B2 JP5937586 B2 JP 5937586B2
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- VBEQCZHXXJYVRD-GACYYNSASA-N uroanthelone Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CS)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)C(C)C)[C@@H](C)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CS)NC(=O)[C@@H](NC(=O)CNC(=O)CNC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CS)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CS)NC(=O)CNC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O)C(C)C)[C@@H](C)CC)C1=CC=C(O)C=C1 VBEQCZHXXJYVRD-GACYYNSASA-N 0.000 description 1
- 230000024883 vasodilation Effects 0.000 description 1
- 229960003726 vasopressin Drugs 0.000 description 1
- 229940118696 vibrio cholerae Drugs 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
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Description
−ある物質と親和性を持つ第1の抗体を基板に付け、
−前記検出される物質をその基板に付けられた前記第1の抗体とある時間接触させ、
−前記抗体と反応しなかった前記物質を洗い流し、
−前記表面に付けられた前記抗体に結合した前記物質を前記物質と親和性を持つ第2の抗体と接触させ、
−前記物質と反応しなかった前記抗体を洗い流し、及び
−前記基板に接合した前記第1の抗体へ結合した前記物質へ結合した前記第2の抗体の存在を検出することを含む。
(a)溶液中に、少なくとも1つの物質を導入し、かつ好ましくは、前記第1の物質と相互作用し得る少なくとも1つの第2の物質を導入し、
(b)ステップ(a)の前記溶液中に、少なくとも2つの磁気粒子又は磁化可能粒子を導入し、前記粒子が前記溶液中に浸漬される表面上に置かれ、
(c)前記粒子が動くように設計される電場、磁場又は電磁場を適用することで、前記物質の相互作用を決定するステップを含み、前記検出される物質間の相互作用は、前記表面上の前記粒子の動きが変化する際に検出される。
これらはまた、人又は動物により次の応答で生成された抗体であり得る:例えばウイルス、菌、プリオン、寄生虫、プロトゾアなどへの感染への応答、同様に、例えば癌、自己免疫疾患例えばバセドウ病、多発性硬化症などの疾患への応答、同様に、例えば農薬、毒物、殺虫剤、除草剤、アレルギー性物質への、中毒、服毒、汚染、薬物注入、摂取、吸入及び注射などへの応答、同様に、例えば骨髄などの移植、例えば腎臓、肺又は肝臓などの器官、手、足、脚部などの手足の移植への応答、さらに同様に人工器官、移植可能な部分、ペースメーカー、人工心臓又は人工股関節の移植への応答である。
有利には、前記第1及び/又は第2の物質の密度が大きいことは、前記第1及び/又は第2の物質が前記容器の底近くに存在することを可能にする。
(a’)前記第1の物質を容器の表面に付ける先行するステップ、
(a’’)前記容器内に溶液を導入するステップ、及び
(a’’’)前記第1の物質と相互作用し得る少なくとも1つの第2の物質を導入するステップを含む。
−前記溶液、
−前記少なくとも1つの粒子、及び
−相互作用可能な前記物質を含む。
この実験では、8つの平底ウェル(参照:MSW002B、 BioFilm Control、 France)の2つのストリップがそれぞれSwH及びSwEとされ使用された。それぞれのストリップで、70μlの抗体溶液がそれぞれのウェルに入れられ、前記溶液は、15μlの、それぞれ、抗ヒトIgG抗体(参照BI2018、 Paris Anticorps、 France)又は抗Ecoli抗体(参照BP2298、 Acris Antibodies、 Germany)のいずれかを、1.5mlのPBS(8 g/lのNaCl(Sigma Aldrich、 USA)、200 mg/lのKCl (Sigma Aldrich、 USA)、1.44g/lのNa2HPO4 (Sigma Aldrich、 USA)、240mg/lのKH2PO4 (Sigma Aldrich、 USA))中で混合して得られたものであり、その後それらを16時間、37℃で安定化させた恒温装置(参照BC240、 Firelabo、 France)中に置いた。
200μlの、ヒト抗FY1抗赤血球細胞抗体を見出すための試薬(IJB−IH Control 3、参照108030357、 Institut Jacques Boy、 France)、2μlの常磁性ミクロビーズ(Ton004N、 BiofilmControl、 France)を含有し、さらに、それぞれ、15μlの注入水又は200μlの抗ヒトIgG抗体(参照BI2018、 Paris Anticorps、 France)又は200μlの抗Ecoli抗体(Reference BP2298、 Acris Antibodies GmbH、 Germany)を含む、それぞれSIg0、SIgH及びSIgEと名付けた溶液を調製した。
この実施例では、3つのストリップで、8つのSBS平坦底ウェルを持つもの(参照MSW002B、 BioFilm Control、 France)、を使用し、それぞれをSwA、SwB及びSwABと指定した。前記ストリップのそれぞれのウェルには、70μlの抗体溶液、それぞれ抗A(参照102010153、 Institut Jacques Boy、 France)、抗B(102010253、 Institut Jacques Boy、 France)、及び抗AB(参照102010353、 Institut Jacques Boy、 France)を入れた。
この実施例では、2つのストリップの8つの平坦底ウェル(参照:MSW002B、 BioFilm Control、 France)が使用された。前記ストリップのそれぞれのウェルに、1.5mlのTS緩衝液(8g/lのNaCl(Sigma Aldrich、 USA)及び1g/lのトリプトン(Difco、 USA))中に15μlの抗ヒトIgG抗体(参照BI2018、 Paris Anticorps、 France)を混合して得られる溶液の50μlを入れた。前記ストリップをその後37℃で安定化された高温装置(参照BC240、 Firelabo、 France)に16時間置いた。
−ウェルF、G及びHのリングと同程度のリングであって、前記ウェルの底部に結合された抗体と赤血球細胞との間の親和結合の不存在による顕著な移動性を示しているか、又は
−拡散したスポットであって、前記ウェルの底部に結合された抗体と赤血球細胞との間の親和結合による前記常磁性ビーズの低減された移動性を示す。
リング及びスポットが、以下表5に示される:
この実施例では、8つの平坦底部ウェルを持つ2つのストリップ(参照:MSW002B、 BioFilm Control、 France)を用いて行い、それぞれSwH及びSwEと指示する。それぞれのストリップで、それぞれのウェルに、それぞれ抗ヒトIgG抗体(参照BI2018、 Paris Anticorps、 France)及び抗Ecoli抗体(参照BP2298、 Acris Antibodies、 Germany)を、1.5mlのPBS(8g/lのNaCl(Sigma Aldrich、 USA)、200mg/lのKCl(Sigma Aldrich、 USA)、1.44g/lのNa2HPO4(Sigma Aldrich、 USA)、 240mg/lのKH2PO4(Sigma Aldrich、 USA))に混合して得られる抗体溶液を入れ、その後それらを37℃で安定化させた高温装置(参照BC240、 Firelabo、 France)に16時間置いた(表6)。
参考文献
1.US 3635678 CLOT−TIMING SYSTEM AND METHOD、 Seitz、 L.J. and Bowen、 J.G.;
2.WO 01/86255、 METHOD AND APPARATUS FOR DETERMINING LOCAL VISCOELASTICITY、 Cronin−Golomb、 M. Shabtai、 Y. Nemet、 B.;
3.EP1544596、 Method and device for determining viscosity、 Kurowski D.; Schoen C.; Peters R.; Bartos H.; Yu Y.;
4.Catterall WA. (2000) Structure and regulation of voltage−gated Ca2+ channels. Annu. Rev. Cell. Dev. Biol. 16:521−55;
5.Gerald Karp、 Biologie cellulaire et moleculaire、 Concepts et experiences [Molecular and Cellular Biology、 Concepts and Experiments]、 2nd edition 2004:
6.Dick GM and Tune JD、 Role of potassium channels in coronary vasodilation、 Exp Biol Med (Maywood). 2010 Jan;235(1):10−22;
7.Mostallino MC et al. Plasticity and function of extrasynaptic GABA(A) receptors during pregnancy and after delivery、 Psychoneuroendocrinology. 2009 Dec; 34 Suppl 1:S74−83.
Claims (12)
- 溶液中の相互作用を検出するための方法であって、
(a)溶液中に、少なくとも1つの第1の物質と、前記第1の物質と相互作用し得る少なくとも1つの第2の物質とを導入するステップ、
(b)ステップ(a)で得られる溶液中に、少なくとも2つの磁気又は磁化可能粒子を導入するステップであり、前記粒子が前記溶液内に浸漬される容器の表面上に存在する、ステップ、
(c)前記粒子が動くように設定される電場、磁場又は電磁場の適用により前記物質間の相互作用を決定するステップであり、前記物質間の相互作用が、前記粒子によって形成されたリングの直径を測定することによって決定される、ステップ、
を含む方法。 - 請求項1に記載の方法であり、前記少なくとも2つの磁気又は磁化可能粒子が、独立して、電荷を帯びた、磁気若しくは磁化可能粒子、又は少なくとも1つの磁気又は磁化可能層でカバーされている粒子である、方法。
- 請求項1又は2のいずれか一項に記載の方法であり、前記少なくとも2つの粒子が、パルス化電磁場にさらされる、方法。
- 請求項1乃至3のいずれか一項に記載の方法であり、前記粒子の移動の加速が、前記電場、磁場又は電磁場の影響下で達成される、方法。
- 請求項1乃至3のいずれか一項に記載の方法であり、前記粒子の移動の減速が、前記電場、磁場又は電磁場の影響下で達成される、方法。
- 請求項1乃至3のいずれか一項に記載の方法であり、前記粒子の軌道の変化が、前記電場、磁場又は電磁場の影響下で達成される、方法。
- 請求項1乃至3のいずれか一項に記載の方法であり、前記相互作用が、前記電場、磁場又は電磁場の影響下での前記粒子のクラスタ化により形成されたリングの直径を測定することによって決定される、方法。
- 請求項1乃至7のいずれか一項に記載の方法であり、前記少なくとも2つの粒子が、動きを検出するために光源によって照射される、方法。
- 請求項1乃至8のいずれか一項に記載の方法であり、前記少なくとも2つの粒子が、シグナル生成粒子である、方法。
- 請求項1乃至9のいずれか一項に記載の方法であり、前記第1の物質が、真核細胞、原核細胞、膜、ウイルス、タンパク質、抗体、抗原、化学分子を含む群から選択される、方法。
- 請求項1乃至9のいずれか一項に記載の方法であり、前記第2の物質が、真核細胞、原核細胞、膜、ウイルス、タンパク質、抗体、抗原、化学分子を含む群から選択される、方法。
- 請求項1乃至11のいずれか一項に記載の方法であり、ステップ(a)の代わりに、先行の
(a’)前記第1の物質を容器の表面に付けるステップ、
(a’’)前記容器に溶液を導入するステップ、及び
(a’’’)前記第1の物質と相互作用し得る少なくとも1つの第2の物質を導入するステップ、
を含む方法。
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FR2866707A1 (fr) * | 2004-02-23 | 2005-08-26 | Thierry Bernardi | Procede et dispositif permettant de detecter la formation et le developpement de biofilms dans un milieu de culture |
FR2883296B1 (fr) * | 2005-03-15 | 2007-05-18 | Nicolas Bara | Procede et dispositif permettant d'isoler les microorganismes |
ATE471516T1 (de) * | 2007-02-23 | 2010-07-15 | Koninkl Philips Electronics Nv | Sensorvorrichtung und verfahren zum detektieren magnetischer partikel |
JP2009069070A (ja) * | 2007-09-14 | 2009-04-02 | Olympus Corp | 標的物質の検出方法及び検出用キット |
WO2009083856A2 (en) * | 2007-12-20 | 2009-07-09 | Koninklijke Philips Electronics N.V. | Concentrated unbound magnetic particle assay for biosensors |
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2010
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Also Published As
Publication number | Publication date |
---|---|
IL223809A (en) | 2017-03-30 |
CA2803743C (fr) | 2018-04-03 |
US20130164736A1 (en) | 2013-06-27 |
MA34341B1 (fr) | 2013-06-01 |
CN103069264B (zh) | 2016-11-02 |
AP3750A (en) | 2016-07-31 |
CA2803743A1 (fr) | 2012-01-05 |
AP2013006701A0 (en) | 2013-01-31 |
MX2013000142A (es) | 2013-06-05 |
JP2013530404A (ja) | 2013-07-25 |
FR2962222A1 (fr) | 2012-01-06 |
EA201291396A1 (ru) | 2013-04-30 |
BR112012033591A2 (pt) | 2016-11-29 |
EA028391B1 (ru) | 2017-11-30 |
EP2588845B1 (fr) | 2016-05-04 |
AU2011273229B2 (en) | 2014-09-18 |
WO2012001312A1 (fr) | 2012-01-05 |
CN103069264A (zh) | 2013-04-24 |
NZ605706A (en) | 2014-07-25 |
ZA201300043B (en) | 2013-09-25 |
AU2011273229A1 (en) | 2013-01-31 |
FR2962222B1 (fr) | 2018-03-30 |
US9746407B2 (en) | 2017-08-29 |
ES2585394T3 (es) | 2016-10-05 |
FR2962221A1 (fr) | 2012-01-06 |
KR20140002597A (ko) | 2014-01-08 |
EP2588845A1 (fr) | 2013-05-08 |
BR112012033591B1 (pt) | 2020-04-14 |
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