公开(公告)号：EP1849003A1

公开(公告)日：2007-10-31

申请号：EP06709506.7

申请日：2006-02-13

申请人： CIS BIO INTERNATIONAL ,  Centre National de la Recherche Scientifique

发明人： ANSANAY, Hervé ,  FINK, Michel ,  MATHIS, Gérard ,  MAUREL, Damien ,  TRINQUET, Eric ,  PIN, Jean-philippe

IPC分类号： G01N33/542 ,  G01N33/74

CPC分类号： G01N33/74 ,  G01N33/542 ,  G01N2333/4719 ,  G01N2333/726 ,  G01N2500/02

摘要： The invention relates to a method for detection of ligands (agonists or antagonists) specific to a receptor (RCPG) coupled to a G protein, comprising the following steps; 1) bring a receptor marked with a member of a donor/acceptor pair into contact with a Gα or Gß? sub-unit of a G protein marked with the other member of the donor/acceptor pair and 2) measure the transfer by proximity effect between the donor and acceptor. The above is of application to the screening of novel medicaments.

公开(公告)号：EP1931997A2

公开(公告)日：2008-06-18

申请号：EP06808261.9

申请日：2006-08-29

申请人： CIS BIO INTERNATIONAL

发明人： TRINQUET, Eric ,  ANSANAY, Hervé ,  MATHIS, Gérard

IPC分类号： G01N33/58

CPC分类号： G01N33/542 ,  G01N33/6872 ,  Y10S435/968

摘要： The invention relates to a method of determining a biological process using a FRET measurement. The inventive method comprises the following steps consisting in: providing a measurement medium containing a lipid membrane with (i) a biological entity X which is coupled to a first member of a pair of FRET partners and (ii) a second biological entity Y which is coupled to the second member of the FRET pair, whereby the energy-donor FRET pair member has a long lifespan and the members of said FRET pair are located on either side of the lipid membrane; exciting the measurement medium at the excitation wavelength of the energy donor member; and measuring the FRET signal or variations in the signal emitted in the culture medium. The invention can be used to detect interactions between biological entities in order to explain biological and pathological processes and to screen novel medicaments.

摘要翻译： 该方法涉及将生物实体（X，Y），加上一对荧光共振能量转移（FRET）的合作伙伴的respectivement成员，在含有脂质膜测量介质：如质膜，其中，所述构件位于两个 膜和一个成员的两侧有很长的寿命。 该膜在在能量供体构件和在培养基中涌出信号FRET信号或变化的激发波长激发的/被测量。

公开(公告)号：EP1817319A2

公开(公告)日：2007-08-15

申请号：EP05824048.2

申请日：2005-12-02

申请人： CIS BIO INTERNATIONAL

发明人： BAZIN, Hervé ,  ANSANAY, Hervé ,  TRINQUET, Eric ,  MATHIS, Gérard

IPC分类号： C07F9/17

CPC分类号： C07F9/6561 ,  C07F9/117 ,  C07F9/5721 ,  C07F9/65517 ,  C07F9/65583

摘要： The invention relates to inositol-phosphate derivatives, in which the inositol-phosphate is substituted by one or two reactive groups G or one or two conjugated molecules M or substances, said reactive group(s) G or said substance(s) or molecule(s) M being linked to IPI via a linkage group L. According to the invention, M is selected from the following group: a tracer, an immunogen, a member of a pair of binding partners, a solid support. The invention is suitable for tools that are used to study the cycle of inositols-phosphates and thus, indirectly, to study seven-domain transmembrane receptors that are coupled to the phospholipase C, receptors having a tyrosine-kinase activity and, generally, enzymes involved in variations in IPI intracellular concentration.

公开(公告)号：EP1817319B1

公开(公告)日：2013-07-03

申请号：EP05824048.2

申请日：2005-12-02

申请人： CIS BIO INTERNATIONAL

发明人： BAZIN, Hervé ,  ANSANAY, Hervé ,  TRINQUET, Eric ,  MATHIS, Gérard

IPC分类号： C07F9/17

CPC分类号： C07F9/6561 ,  C07F9/117 ,  C07F9/5721 ,  C07F9/65517 ,  C07F9/65583

公开(公告)号：EP2329272A2

公开(公告)日：2011-06-08

申请号：EP09740420.6

申请日：2009-07-30

申请人： Cis-Bio International

发明人： ZWIER, Jurriaan ,  POOLE, Robert ,  ANSANAY, Hervé ,  FINK, Michel ,  TRINQUET, Eric

IPC分类号： G01N33/566 ,  G01N33/58

CPC分类号： G01N33/542 ,  G01N33/5035 ,  G01N33/58 ,  G01N2458/40 ,  Y10T436/13

摘要： A method for detecting the internalization of a transmembrane protein of interest expressed at the surface of a cell, comprising the following steps: a) labelling the protein of interest with a fluorescent metal complex, the lifetime of which is greater than 0.1 ms; b) adding to the reaction medium a composition capable of causing the internalization of the protein of interest; c) adding to the reaction medium a modulating agent selected from: a. a fluorescent or nonfluorescent FRET acceptor compound compatible with said fluorescent metal complex, the final concentration of which in the reaction medium is greater than 10
‑7 M; b. a reducing agent, the redox potential of which is less than +0.1 V and preferably between 0.25 and 0.75 V; c. an agent which binds specifically, by noncovalent bonding, with the fluorescent metal complex; d. a metal ion which competes with the rare earth so as to form a nonfluorescent metal complex; d) measuring the luminescence emitted by the reaction medium at the emission wavelength of the fluorescent metal complex and/or at the emission wavelength of the modulating compound when said compound is a fluorescent acceptor compound; e) comparing the signal measured in step d) with a reference signal measured on cells having been subjected only to steps a) and c). Use: Method for detecting membrane protein internalization.

摘要翻译： 一种检测细胞表面表达的感兴趣的跨膜蛋白的内化的方法，包括以下步骤：a）用荧光金属配合物标记感兴趣的蛋白质，其寿命大于0.1ms; b）向反应介质中加入能够引起感兴趣蛋白质内化的组合物; c）向反应介质中加入选自以下的调节剂：a。 与所述荧光金属配合物相容的荧光或非荧光FRET受体化合物，其在反应介质中的最终浓度大于10 -7 M; 湾 还原剂，其氧化还原电位低于+ 0.1V，优选在0.25和0.75V之间; C。 通过非共价键特异性结合荧光金属配合物的试剂; d。 与稀土竞争以形成无荧光金属络合物的金属离子; d）当所述化合物是荧光受体化合物时，在荧光金属络合物的发射波长处和/或在调制化合物的发射波长处测量由反应介质发射的发光; e）将步骤d）中测得的信号与仅在经过步骤a）和c）的细胞上测量的参考信号进行比较。 用途：检测膜蛋白内在化的方法。