Fluorescent intensity assay for protein or peptide binding...

C - Chemistry – Metallurgy – 12 – Q

Patent

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C12Q 1/68 (2006.01) G01N 33/53 (2006.01) G01N 33/542 (2006.01) G01N 33/58 (2006.01) G01N 33/68 (2006.01)

Patent

CA 2358065

A method for assaying binding between a fluorophore-labeled compound and an unlabeled compound is provided. The method includes detecting a quenching effect on fluorescence emitted by the fluorophore-labeled compound resulting from binding. The binding is specific and other than nucleobase to nucleobase. The method is conducted without separating complexes of the fluorophore-labeled compound and the unlabeled compound from the fluorophore-labeled compound prior to quenching effect detecting, and without providing a signal quenching agent to quench fluorescent light. Preferably, the fluorophore-labeled compound is a nucleic acid and the unlabeled compound is a protein. The method can be used for a variety of applications, including screening for drug candidates having optimum binding properties, and quantifying the binding affinity of DNA binding proteins for nucleic acids.

L'invention concerne un procédé de dosage de la liaison entre un composé marqué fluorophore et un composé non marqué. Le procédé consiste à détecter un effet de réduction sur la fluorescence émise par le composé marqué fluorophore résultant de la liaison. La liaison est spécifique et autre que nucléobase à nucléobase. Le procédé est exécuté sans que le composé marqué fluorophore soit séparé des complexes du composé marqué fluorophore et du composé non marqué avant la détection de l'effet de réduction, et sans présentation d'un agent de réduction de signal pour réduire la lumière fluorescente. De préférence, le composé marqué fluorophore est un acide nucléique et le composé non marqué une protéine. Le procédé peut servir pour diverses applications, notamment le criblage de médicaments candidats possédant des propriétés de liaison optimales, et la quantification de l'affinité de liaison de protéines de liaison d'ADN pour des acides nucléiques.

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