Method of measuring enzyme activity and method of evaluating...

C - Chemistry – Metallurgy – 12 – Q

Patent

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C12Q 1/25 (2006.01) C12Q 1/48 (2006.01) C12Q 1/527 (2006.01) G01N 37/00 (2006.01)

Patent

CA 2602666

It is intended to provide a method of measuring an activity, which is a method of measuring an activity of an enzymatic reaction in which a lipid-soluble reaction product is formed by catalyzing an addition reaction of two or more kinds of substrates, characterized by comprising a labeling step in which any one of the substrates is labeled, a reaction step in which an enzymatic reaction is carried out in the presence of an enzyme, all substrates and an acceptor, and a detection step in which the reaction product is detected by bringing the labeled molecule and a molecule to be detected close to each other via the acceptor and transferring an energy generated by the labeled molecule to the molecule to be detected, and a method of evaluating a compound utilizing the method of measuring an activity. By the methods of the invention, it is possible to perform a treatment from the enzymatic reaction to the measurement of an enzyme activity with the simple steps and it is possible to provide a method of measuring an enzyme activity which can be applied to an HTS evaluation system.

L'invention concerne un procédé permettant de mesurer l'activité d'une réaction enzymatique, dans laquelle un produit de réaction soluble dans un liquide est formé par catalyse d'une réaction d'addition de deux ou davantage de substrats. Le procédé est caractérisé par les étapes consistant à: marquer l'un des substrats; mettre en oeuvre une réaction enzymatique en présence d'une enzyme, tous les substrats et un accepteur; et détecter le produit de réaction en mettant la molécule marquée à proximité étroite d'une molécule à détecter, par l'intermédiaire de l'accepteur; et transférer l'énergie produite par la molécule marquée vers la molécule à détecter. L'invention concerne aussi un procédé d'évaluation d'un composé utilisant ce procédé de mesure de l'activité enzymatique. Les procédés de l'invention permettent, par l'exécution d'étapes simples, de mettre en oeuvre une réaction enzymatique et de mesurer l'activité enzymatique. Le procédé de mesure de l'activité enzymatique peut être appliqué dans un système d'évaluation par criblage à haut débit (HTS).

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