Multiple enzyme assays

Details Multiple enzyme assays Multiple enzyme assays

C - Chemistry, Metallurgy

12

Q

C12Q 1/66 (2006.01) C12Q 1/34 (2006.01) C12Q 1/37 (2006.01) C12Q 1/42 (2006.01) C12Q 1/44 (2006.01) G01N 21/00 (2006.01) C12N 9/02 (2006.01) C12N 9/16 (2006.01) C12N 9/24 (2006.01) C12N 9/38 (2006.01) C12N 9/48 (2006.01)

Patent

CA 2353872

The present invention discloses multiple enzyme assays which measure the activity of at least one endogenous enzyme in a single aliquot and a method of measuring the activity of multiple enzymes in an aliquot of a sample extract, wherein at least one of the enzymes is an endogenous enzyme. In one embodiment of the invention the activity of a first enzyme is quantified by measuring the light signal produced by degradation of a first enzyme substrate by the first enzyme and the activity of the second enzyme is quantified by measuring the light signal produced by the degradation of a second substrate. In the method of the present invention, both quantifications are performed on the same aliquot of sample extract. Different embodiments of the present invention provide for the detection of more than one endogenous enzyme and for the detection of at least one reporter enzyme and at least one endogenous enzyme. The present invention also discloses kits for detecting the activity of multiple enzymes.

L'invention concerne des dosages multi-enzymatiques permettant de mesurer l'activité d'au moins une enzyme endogène, dans une seule aliquote, ainsi qu'un procédé de mesure de l'activité multi-enzymatique dans une partie aliquote d'un extrait d'échantillon, l'une de ces enzymes au moins étant une enzyme endogène. Dans un mode de réalisation de l'invention, on quantifie l'activité d'une première enzyme en mesurant le signal lumineux produit par la dégradation d'un premier substrat enzymatique et on quantifie l'activité d'une seconde enzyme en mesurant le signal lumineux produit par la dégradation d'un second substrat. Dans le procédé de l'invention, on exécute les deux quantifications sur la même aliquote d'un extrait d'échantillon. Différents modes de réalisation de l'invention permettent de détecter plus d'une enzyme endogène, et de détecter au moins une enzyme rapporteur et au moins une enzyme endogène. L'invention concerne également des trousses de détection de l'activité multi-enzymatique.

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