Fluorescent intensity method for assaying binding between...

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G01N 33/542 (2006.01)

Patent

CA 2377815

A method for assaying specific binding between a fluorophore-labeled probe and an unlabeled target is provided. The method includes detecting a quenching effect on fluorescence emitted by the fluorophore-labeled probe resulting from binding. The method is conducted without separating complexes of the target and probe from the free target and free probe prior to quenching effect detecting, and without providing a signal quenching agent to quench fluorescent light. Preferably, the probe and target are amino acid-containing compounds, such as proteins. The method can be used for a variety of applications, including screening for drug candidates having optimum binding properties, and quantifying and classifying the binding characteristics between peptide- containing compounds. The method is more sensitive than conventional assays, enabling the analysis of minute samples and low affinity binding interactions between receptors and ligands that are below the detection limits of conventional technology.

L'invention concerne un procédé permettant de déterminer une liaison spécifique entre une sonde marquée fluorophore et une cible non marquée. Ce procédé consiste à détecter un effet de réduction sur une fluorescence émise par la sonde marquée fluorophore résultant de ladite liaison. Ce procédé est mis en oeuvre sans séparer les complexes de la cible et de la sonde, de la cible et de la sonde libres avant la détection de l'effet de réduction, et sans fournir d'agent de réduction de signal, afin de réduire la lumière fluorescente. La sonde et la cible sont, de préférence, des composés contenant un acide aminé, tels que des protéines. On peut utiliser ce procédé dans une variété d'applications, notamment le criblage de médicaments candidats, la quantification, et la classification de caractéristiques de liaison entre des composés contenant des peptides. Ce procédé plus sensible que les analyses conventionnelles, permet d'analyser des échantillons minuscules et des interactions de liaisons à faible affinité entre des récepteurs et des ligands, qui se trouvent en-dessous des limites de détection des technologies conventionnelles.

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